E-selectin antagonists

ABSTRACT

Compounds, compositions and methods are provided for inhibiting in vitro and in vivo processes mediated by E-selectin binding. More specifically, particular glycomimetic compounds are described, wherein the compounds are E-selectin antagonists.

This application claims the benefit under 35 U.S.C. §119(e) of U.S.Provisional Patent Application No. 61/382,716 filed Sep. 14, 2010, wherethis provisional application is incorporated herein by reference in itsentirety.

BACKGROUND

1. Technical Field

The present invention relates generally to compounds, compositions andmethods for inhibiting E-selectin, and more specifically to particularglycomimetics which are E-selectin antagonists.

2. Description of the Related Art

When a tissue is infected or damaged, the inflammatory process directsleukocytes and other immune system components to the site of infectionor injury. Within this process, leukocytes play an important role in theengulfment and digestion of microorganisms. Thus, the recruitment ofleukocytes to infected or damaged tissue is critical for mounting aneffective immune defense.

Selectins are a group of structurally similar cell surface receptorsthat are important for mediating leukocyte binding to endothelial cells.These proteins are type 1 membrane proteins and are composed of an aminoterminal lectin domain, an epidermal growth factor (EGF)-like domain, avariable number of complement receptor related repeats, a hydrophobicdomain spanning region and a cytoplasmic domain. The bindinginteractions appear to be mediated by contact of the lectin domain ofthe selectins and various carbohydrate ligands.

There are three known selectins: E-selectin, P-selectin and L-selectin.E-selectin is found on the surface of activated endothelial cells, whichline the interior wall of capillaries. E-selectin binds to thecarbohydrate sialyl-Lewis^(x) (SLe^(x)), which is presented as aglycoprotein or glycolipid on the surface of certain leukocytes(monocytes and neutrophils) and helps these cells adhere to capillarywalls in areas where surrounding tissue is infected or damaged; andE-selectin also binds to sialyl-Lewis^(a) (SLe^(a)), which is expressedon many tumor cells. P-selectin is expressed on inflamed endothelium andplatelets, and also recognizes SLe^(x) and SLe^(a), but also contains asecond site that interacts with sulfated tyrosine. The expression ofE-selectin and P-selectin is generally increased when the tissueadjacent to a capillary is infected or damaged. L-selectin is expressedon leukocytes. Selectin-mediated intercellular adhesion is an example ofa selectin-mediated function.

Modulators of selectin-mediated function include the PSGL-1 protein (andsmaller peptide fragments), fucoidan, glycyrrhizin (and derivatives),anti-selectin antibodies, sulfated lactose derivatives, and heparin. Allhave shown to be unsuitable for drug development due to insufficientactivity, toxicity, lack of specificity, poor ADME characteristicsand/or availability of material.

Although selectin-mediated cell adhesion is required for fightinginfection and destroying foreign material, there are situations in whichsuch cell adhesion is undesirable or excessive, resulting in tissuedamage instead of repair. For example, many pathologies (such asautoimmune and inflammatory diseases, shock and reperfusion injuries)involve abnormal adhesion of white blood cells. Such abnormal celladhesion may also play a role in transplant and graft rejection. Inaddition, some circulating cancer cells appear to take advantage of theinflammatory mechanism to bind to activated endothelium. In suchcircumstances, modulation of selectin-mediated intercellular adhesionmay be desirable.

Accordingly, there is a need in the art for identifying inhibitors ofselectin-mediated function, e.g., of selectin-dependent cell adhesion,and for the development of methods employing such compounds to inhibitconditions associated with excessive selectin activity. The presentinvention fulfills these needs and further provides other relatedadvantages.

BRIEF SUMMARY

Briefly stated, this invention provides compounds, compositions andmethods for inhibiting E-selectin. The compounds are particularE-selectin antagonists. Such compounds may be combined with apharmaceutically acceptable carrier or diluent to form a pharmaceuticalcomposition. The compounds or compositions may be used in a method toinhibit an E-selectin-mediated function, such as inhibiting anE-selectin-mediated intercellular adhesion.

In one aspect of the present invention, a compound is provided havingone of the following formulae:

A compound of the present invention includes physiologically acceptablesalts thereof. A compound of the present invention in combination with apharmaceutically acceptable carrier or diluent provides a composition ofthe present invention. In the chemical formula herein, the abbreviation“Me” or a single line extending from a carbon implied by theintersection of two other lines, represents a methyl group.

These and other aspects of the present invention will become apparentupon reference to the following detailed description and attacheddrawings. All references disclosed herein are hereby incorporated byreference in their entirety as if each was incorporated individually.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a diagram illustrating the synthesis of the precursorglycomimetic ligand 1. Glycomimetic ligand 1 was synthesized accordingto International Application Publication No. WO 2008/060378 (e.g.,Example 1 and FIG. 1).

FIG. 2 is a diagram of a general description of the synthesis ofselectin antagonists with triazole linker.

FIG. 3 is a diagram illustrating the synthesis of the alkynylglycomimetic ligands.

FIG. 4 is a diagram illustrating the synthesis of the indole derivativesstarting from 5-nitro-1H-indole.

FIG. 5 is a diagram illustrating the synthesis of the indole derivativesstarting from 2-iodo-4-nitroaniline (Larock indole synthesis).

FIG. 6 is a diagram illustrating the synthesis of the selectinantagonists with triazole linker.

FIG. 7 is a diagram illustrating the synthesis of the selectinantagonists with piperazine linker.

DETAILED DESCRIPTION

As noted above, the present invention provides compounds which areE-selectin antagonists, compositions thereof and methods for inhibitingE-selectin-mediated functions. Such antagonists may be used in vitro orin vivo, to inhibit E-selectin-mediated functions in a variety ofcontexts. Examples of E-selectin-mediated functions includeintracellular adhesion.

The E-selectin antagonists of the present invention are particularcompounds that surprisingly show significant inhibitory activity atnanomolar concentrations. The compounds include physiologicallyacceptable salts thereof. The compounds have the following formulae:

Binding affinities of the compounds are set forth in Table 1 below.

All compounds of the present invention or useful thereto (e.g., forpharmaceutical compositions or methods of treating), includephysiologically acceptable salts thereof. Examples of such salts are Na,K, Li, Mg, Ca and Cl.

Compounds as described herein may be present within a pharmaceuticalcomposition. A pharmaceutical composition comprises one or morecompounds in combination with (i.e., not covalently bonded to) one ormore pharmaceutically or physiologically acceptable carriers, diluentsor excipients. Such compositions may comprise buffers (e.g., neutralbuffered saline or phosphate buffered saline), carbohydrates (e.g.,glucose, mannose, sucrose or dextrans), mannitol, proteins, polypeptidesor amino acids such as glycine, antioxidants, chelating agents such asEDTA or glutathione, adjuvants (e.g., aluminum hydroxide) and/orpreservatives. Within yet other embodiments, compositions of the presentinvention may be formulated as a lyophilizate. Compositions of thepresent invention may be formulated for any appropriate manner ofadministration, including for example, topical, oral, nasal,intravenous, intracranial, intraperitoneal, subcutaneous, orintramuscular administration.

The compositions described herein may be administered as part of asustained release formulation (i.e., a formulation such as a capsule orsponge that effects a slow release of compound followingadministration). Such formulations may generally be prepared using wellknown technology and administered by, for example, oral, rectal orsubcutaneous implantation, or by implantation at the desired targetsite. Carriers for use within such formulations are biocompatible, andmay also be biodegradable; preferably the formulation provides arelatively constant level of compound release. The amount of compoundcontained within a sustained release formulation depends upon the siteof implantation, the rate and expected duration of release and thenature of the condition to be treated.

The above described compounds including equivalents thereof are usefulin methods of the present invention to treat individuals in needthereof. As used herein, such individuals include humans, as well asnon-human warm-blooded animals such as non-human mammals. A preferredindividual for treatment is a human. Typically a compound will beadministered to an individual as a pharmaceutical composition, i.e., incombination with a pharmaceutically acceptable carrier or diluent.

The above described compounds may be administered in a mannerappropriate to the disease to be treated. Appropriate dosages and asuitable duration and frequency of administration may be determined bysuch factors as the condition of the patient, the type and severity ofthe patient's disease and the method of administration. In general, anappropriate dosage and treatment regimen provides the compound(s) in anamount sufficient to provide therapeutic and/or prophylactic benefit.

Within particularly preferred embodiments of the invention, a compoundmay be administered at a dosage ranging from 0.001 to 1000 mg/kg bodyweight (more typically 0.01 to 1000 mg/kg), on a regimen of single ormultiple daily doses. Appropriate dosages may generally be determinedusing experimental models and/or clinical trials. In general, the use ofthe minimum dosage that is sufficient to provide effective therapy ispreferred. Patients may generally be monitored for therapeuticeffectiveness using assays suitable for the condition being treated,which will be familiar to those of ordinary skill in the art.

In an embodiment, an individual who is in need of treatment of a disease(or complication associated therewith) in which an E-selectin-mediatedfunction is involved is administered at least one (i.e., one or more) ofthe above described compounds in an amount effective for the treatment.

A variety of conditions are associated with a selectin-mediatedfunction. Such conditions include, for example, tissue transplantrejection, platelet-mediated diseases (e.g., atherosclerosis andclotting), hyperactive coronary circulation, acute leukocyte-mediatedlung injury (e.g., adult respiratory distress syndrome (ARDS)), Crohn'sdisease, inflammatory diseases (e.g., inflammatory bowel disease),autoimmune diseases (MS, myasthenia gravis), infection, cancer (andmetastasis), thrombosis, wounds (and wound-associated sepsis), burns,spinal cord damage, digestive tract mucous membrane disorders(gastritis, ulcers), osteoporosis, rheumatoid arthritis, osteoarthritis,asthma, allergy, psoriasis, septic shock, traumatic shock, stroke,nephritis, atopic dermatitis, frostbite injury, adult dyspnoea syndrome,ulcerative colitis, systemic lupus erythematosus, diabetes andreperfusion injury following ischaemic episodes. Compounds may also beadministered to a patient prior to heart surgery to enhance recovery.Other uses include pain management, prevention of restinosis associatedwith vascular stenting, and for undesirable angiogenesis, e.g.,associated with cancer.

The term “treatment,” as set forth above, refers to any of a variety ofpositive effects from the treatment including, for example, eradicatinga complication associated with the disease, relieving to some extent acomplication, slowing or stopping progression of the disease, andprolonging the survival time of the recipient. The treatment may be usedin conjunction with one or more other therapies for any of the illnesses(or complications associated therewith) described above.

Compounds as described above are capable, for example, of inhibitingselectin-mediated cell adhesion. This ability may generally be evaluatedusing any of a variety of in vitro assays designed to measure the effecton adhesion between selectin-expressing cells (e.g., adhesion betweenleukocytes or tumor cells and platelets or endothelial cells). Forexample, such cells may be plated under standard conditions that, in theabsence of compound, permit cell adhesion. In general, a compound is aninhibitor of selectin-mediated cell adhesion if contact of the testcells with the compound results in a discernible inhibition of celladhesion. For example, in the presence of compounds (e.g., micromolarlevels), disruption of adhesion between leukocytes or tumor cells andplatelets or endothelial cells may be determined visually withinapproximately several minutes, by observing the reduction of cellsinteracting with one another.

The following Examples are offered by way of illustration and not by wayof limitation.

EXAMPLES Example 1 Synthesis of the Alkynyl Glycomimetic Ligands (FIG.3) Synthesis of Intermediate 2 6-(Trimethylsilyl)-hex-5-yn-1-ol

Under argon, hex-5-yn-1-ol (1.03 g, 10.5 mmol) was dissolved inanhydrous THF (10 mL) and cooled to −90° C. ^(t)BuLi (1.6 M in pentane;14.4 mL, 23.1 mmol) was added over a period of 10 minutes, which lead torefreezing of the reaction mixture. Gradual warming to −10° C. over 15min led to liquefaction, and after 2 h of stirring, Me₃SiCl (3.0 mL,23.1 mmol) was added. After 2 h, 1 M aq. HCl (3 mL) was added andstirring was continued for another hour. The reaction mixture wasextracted with Et₂O (3×40 mL), the combined organic layers were washedwith satd. aq. NaHCO₃ (20 mL) and brine (20 mL). The dried (Na₂SO₄)organic phases were concentrated in vacuo, and alcohol 2 was obtained asa colorless oil (2.10 g), which was used without further purification.

Synthesis of Intermediate 3 3-(Trimethylsilyl)prop-2-ynylmethanesulfonate

3-(trimethylsilyl)prop-2-yn-1-ol (1.56 g, 12.2 mmol) was dissolved inanhydrous CH₂Cl₂ (37 mL) under argon. The solution was cooled to −78°C., followed by addition of Et₃N (3.37 mL, 24.3 mmol). MeSO₂Cl (0.948mL, 12.2 mmol) was added to the solution over a period of 10 min,leading to the formation of a white precipitate. After stirring at −78°C. for 30 min, the reaction mixture was washed with 0.5 M aq. HCl (15mL), satd. aq. NaHCO₃ (20 mL), and brine (20 mL). After extraction ofthe aqueous layers with CH₂Cl₂ (2×30 mL), the combined organic layerswere dried over Na₂SO₄, and the solvent was removed in vacuo to give themesylate 3 as a pale yellow liquid (2.46 g) which was used withoutfurther purification.

Synthesis of Intermediate 4 4-(Trimethylsilyl)but-3-ynylmethanesulfonate

Following the procedure for 3, MeSO₂Cl (2.05 mL, 26.3 mmol) was added toa solution of 4-(trimethylsilyl)but-3-yn-1-ol (3.12 g, 21.9 mmol) andEt₃N (6.07 mL, 43.8 mmol) in anhydrous CH₂Cl₂ (65 mL) at −15° C. After1.5 h, the reaction mixture was worked up, and the mesylate 4 wasobtained as a pale yellow oil (4.88 g), which was used without furtherpurification.

Synthesis of Intermediate 5 5-(Trimethylsilyl)pent-4-ynylmethanesulfonate

Following the procedure for 3, MeSO₂Cl (1.37 mL, 17.6 mmol) was added toa solution of 5-(trimethylsilyl)pent-4-yn-1-ol (2.29 g, 14.7 mmol) andEt₃N (4.07 mL, 29.4 mmol) in anhydrous CH₂Cl₂ (50 mL) at −15° C. After 2h, the reaction mixture was worked up, and the mesylate 5 was obtainedas a pale yellow oil (3.56 g), which was used without furtherpurification.

Synthesis of Intermediate 6 6-(Trimethylsilyl)hex-5-ynylmethanesulfonate

MeSO₂Cl (0.98 mL, 12.6 mmol) was added to a solution of 2 (2.10 g) andEt₃N (2.9 mL, 21.0 mmol) in anhydrous CH₂Cl₂ (35 mL) at −15° C. Thereaction mixture was allowed to warm to r.t., and after 1.5 h, it waswashed with 0.5 M aq. HCl (10 mL), satd. aq. NaHCO₃ (15 mL), and brine(15 mL). After extraction of the aqueous layers with CH₂Cl₂ (2×30 mL),the combined organic layers were dried over Na₂SO₄, and the solvent wasremoved in vacuo. The residue was purified by silica gel chromatography(petrol ether/CH₂Cl₂ linear gradient) to afford mesylate 6 as acolorless liquid (2.26 g).

Synthesis of Intermediate 7 (3-Azidoprop-1-ynyl)trimethylsilane

Mesylate 3 (2.46 g, 11.9 mmol) was dissolved in anhydrous DMF (50 mL)under argon. NaN₃ (0.87 g, 13.4 mmol) was added, and the suspension washeated to 65° C. After vigorous stirring of the viscous reaction mixturefor 35 min, water (30 mL) was added, and the suspension was filteredthrough a plug of celite. The celite was washed with diethyl ether (80mL). The water-DMF mixture was extracted with Et₂O (3×60 mL), and theorganic layers were thoroughly washed with water (2×40 mL) and brine (40mL). The combined organic layers were dried over Na₂SO₄, the solvent wasremoved in vacuo, and azide 7 was obtained as a yellow liquid (1.82 g),which was used without further purification.

Synthesis of Intermediate 8 (4-Azidobut-1-ynyl)trimethylsilane

Following the procedure for 7, mesylate 4 (4.88 g, 22.1 mmol) wasreacted with NaN₃ (1.57 g, 24.1 mmol) in anhydrous DMF (55 mL). After 3h, the reaction was filtered and worked up to give azide 8 as a yellowliquid (3.35 g), which was used without further purification.

Synthesis of Intermediate 9 (5-Azidopent-1-ynyl)trimethylsilane

Following the procedure for 7, mesylate 5 (3.56 g, 15.2 mmol) wasreacted with NaN₃ (1.10 g, 16.9 mmol) in anhydrous DMF (35 mL). After2.5 h, the reaction was filtered and worked up to give azide 9 as ayellow liquid (2.62 g), which was used without further purification.

Synthesis of Intermediate 10 (5-Azidohex-1-ynyl)trimethylsilane

Following the procedure for 7, mesylate 6 (2.26 g, 9.10 mmol) wasreacted with NaN₃ (716 mg, 11.0 mmol) in anhydrous DMF (20 mL). After 1h, the reaction was filtered and worked up to afford azide 10 as ayellow liquid (1.41 g), which was used without further purification.

¹H NMR (500 MHz, CDCl₃): δ 0.12 (s, 9H, Me₃Si), 1.57-1.66 (m, 2H, H-3),1.81-1.89 (m, 2H, H-2), 2.99 (s, 3H, MeSO₂), 4.24 (t, J=6.4 Hz, 2H,H-1); ¹³C NMR (125 MHz, CDCl₃): δ 0.18 (Me₃Si), 19.32 (C-4), 24.49(C-3), 28.22 (C-2), 37.49 (MeSO₂), 69.59 (C-1), 85.52 (C-6), 106.19(C-5).

Synthesis of Intermediate 11 3-(Trimethylsilyl)-prop-2-yn-1-amine

To a solution of azide 7 (1.82 g, 11.9 mmol) in THF (50 mL) and water (1mL) was added PPh₃ (3.18 g, 12.2 mmol). The solution was stirred at 50°C. for 2 h. After careful removal of the solvent in vacuo (≧200 mbar),the reaction mixture was subjected to vacuum distillation (oil bathtemperature: 100° C.; approx. 5 mbar; additional cooling trap withliquid nitrogen) to give amine 11 as a colorless liquid (1.29 g, 83%over 3 steps).

¹H-NMR (500 MHz, CDCl₃): δ 0.11 (s, 9H, TMS-H), 1.37 (s, 2H, NH₂), 3.38(s, 2H, H-1). ¹³C-NMR (125 MHz, CDCl₃): δ 0.09 (3C, TMS-C), 32.51 (C-1),86.72 (C-2), 107.41 (C-3); IR (film on NaCl): ν=3375 (s, NH₂), 3295 (s,NH₂), 3181 (m, NH₂), 2960 (vs), 2900 (s), 2850 (m), 2167 (vs, C≡C), 1599(w, NH₂), 1409 (w), 1379 (w), 1332 (s), 1251 (s, SiCH₃), 1074 (m), 998(s), 955 (m), 908 (m), 842 (m, Me₃Si), 760 (m, Me₃Si), 734 (m), 699 (m),642 (m), 584 (m) cm⁻¹.

The NMR spectral data were in accordance with previously publishedvalues [I. MacInnes, J. C. Walton, J. Chem. Soc., Perkin Trans. 2 1987,8, 1077-1082].

Synthesis of Intermediate 12 4-(Trimethylsilyl)-but-3-yn-1-amine

To a solution of azide 8 (3.35 g, 20.0 mmol) in THF (90 mL) and water(1.8 mL) was added PPh₃ (5.76 g, 21.9 mmol). The solution was stirred at50° C. for 1.5 h. After removal of the solvent in vacuo, the reactionmixture was subjected to vacuum distillation (oil bath temperature: 120°C.; approx. 0.1 mbar; additional cooling trap with liquid nitrogen) togive the amine 12 as a colorless liquid (2.49 g, 80% over 3 steps).

¹H NMR (500 MHz, CDCl₃): δ 0.12 (s, 9H, Me₃Si), 1.40 (s, 2H, NH₂), 2.34(t, 2H, H-1), 2.80 (t, 2H, H-2); ¹³C NMR (125 MHz, CDCl₃): δ 0.23(Me₃Si), 25.08 (C-2), 41.09 (C-1), 86.29 (C-3), 105.09 (C-4). IR (filmon NaCl): ν3373 (m, NH₂), 3297 (m, NH₂), 2959 (vs), 2896 (s,), 2868 (s,C—H), 2174 (vs, C≡C), 1678 (m, NH₂), 1621 (w), 1455 (m), 1425 (m), 1409(m), 1383 (m), 1355 (m), 1250 (vs, SiCH₃), 1179 (vw), 1157 (vw), 1119(vw), 1085 (w), 1034 (s), 979 (m), 908 (s), 844 (vs, Me₃Si), 760 (vs,Me₃Si), 734 (vs), 699 (m), 639 (s), 574 (m) cm⁻¹.

The ¹H NMR spectral data were in accordance with previously publishedvalues [M. Pullagurla, M. Dukat, B. L. Roth, V. Setola, B. A. Glennon,Med. Chem. Res. 2005, 14, 1-18.].

Synthesis of Intermediate 13 5-(Trimethylsilyl)-pent-4-yn-1-amine

To a solution of azide 9 (1.30 g, 7.17 mmol) in THF (100 mL) and water(1 mL) was added PPh₃ (2.24 g, 8.5 mmol). The solution was stirred at50° C. for 4.5 h. After removal of the solvent in vacuo, the reactionmixture was subjected to vacuum distillation (oil bath temperature: 120°C.; approx. 0.1 mbar) to give amine 13 as a colorless oil (0.973 g, 43%over 3 steps).

¹H NMR (500 MHz, CDCl₃): δ 0.12 (s, 9H, Me₃Si), 1.18 (s, 2H, NH₂), 1.62(p, 2H, H-2), 2.27 (t, J=7.0 Hz, 2H, H-3), 2.77 (t, J=6.9 Hz, 2H, H-1);¹³C NMR (125 MHz, CDCl₃): δ 0.23 (Me₃Si), 17.42 (C-3), 32.42 (C-2),41.34 (C-1), 84.95 (C-4), 106.92 (C-5). IR (film on NaCl): ν3376 (m,NH₂), 3297 (w, NH₂), 3181 (w, NH₂), 2955 (vs), 2861 (vs), 2173 (vs,C≡C), 1682 (w), 1602 (w, NH₂), 1431 (s), 1408 (m), 1324 (m), 1250 (vs,Me₃Si), 1087 (s), 1000 (m), 914 (s), 843 (vs, Me₃Si), 760 (vs, Me₃Si),734 (vs), 698 (s), 640 (s), 579 (w) cm⁻¹.

The spectral data were in accordance with previously published values[Y. W. Li, T. J. Marks, J. Am. Chem. Soc. 1996, 118, 9295-9306.], [Y.Li, P.-F. Fu, T. J. Marks, Organometallics 1994, 13, 439-440.]

Synthesis of Intermediate 14 6-(Trimethylsilyl)-hex-5-yn-1-amine

To a solution of azide 10 (1.41 g, 7.22 mmol) in THF (100 mL) and water(1 mL) was added PPh₃ (2.27 g, 8.65 mmol). The solution was stirred at50 to 60° C. for 4.5 h. After removal of the solvent in vacuo, thereaction mixture was subjected to vacuum distillation (oil bathtemperature: 120° C.; approx. 0.1 mbar) to give the amine 14 as acolorless oil (1.10 g, 62% over 4 steps).

¹H NMR (500 MHz, CDCl₃): δ 0.43 (s, 9H, Me₃Si), 1.39 (s, 2H, NH₂),1.82-1.87 (m, 4H, H-2, H-3), 2.53 (t, J=6.6 Hz, 2H, H-4), 2.98-3.03 (m,2H, H-1); ¹³C NMR (125 MHz, CDCl₃): δ 0.06 (Me₃Si), 19.62 (C-4), 25.89(C-3), 32.87 (C-2), 41.65 (C-1), 84.89 (C-6), 107.12 (C-5). IR: ν3371(s, NH₂), 3296 (s, NH₂), 3181 (m, NH₂), 2957 (vs), 2929 (vs), 2901 (vs),2860 (vs), 2173 (vs, C≡C), 1596 (m, NH₂), 1455 (m), 1431 (m), 1408 (m),1364 (w), 1326 (m), 1249 (vs, Me₃Si), 1051 (m), 1027 (m), 998 (w), 937(m), 842 (vs, Me₃Si), 760 (vs, Me₃Si), 698 (s), 639 (s), 619 (vw), 461(s) cm⁻¹.

Synthesis of Intermediate 15(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-cyclohexanecarboxylicacid

Ester 1 (268 mg, 0.355 mmol) was dissolved in a 1 M solution of NaOH inH₂O/MeOH 1:1 (8 mL) and stirred at r.t. for 3 h. The reaction wasstopped by addition of 0.5 M aq. HCl (1.6 mL), and the solvent wasremoved in vacuo. The residue was purified using preparative HPLC-MS(H₂O/MeCN+0.2% HCOOH) yielding diacid 15 as a white powder (207 mg,79%). The starting material was partially recovered (25 mg, 9%).

¹H-NMR (500 MHz, MeOD): δ 0.50-0.75 (m, 4H, Cy), 0.86-0.97 (m, 1H, Cy),1.11 (d, J=6.4 Hz, 3H, Me), 1.06-1.16 (m, 2H, H-6_(a), H-2_(a)), 1.33(d, J=6.5 Hz, 3H, Fuc H-6), 1.19-1.38 (m, 5H, 5Cy), 1.38-1.47 (m, 1H,Lac H-3_(a)), 1.47-1.56 (m, 1H, Lac H-3_(b)), 1.56-1.72 (m, 2H, Cy,H-5), 1.78 (d, J=12.8 Hz, 1H, H-6), 2.29-2.40 (m, 2H, H-2, H-1), 3.11(t, J=9.5 Hz, 1H, H-4), 3.58 (t, J=5.6 Hz, 1H, Gal H-5), 3.64-3.80 (m,6H, Gal H-3, H-3, Fuc H-2, 2Gal H-6, Fuc H-4), 3.85 (dd, J=3.0, 10.3 Hz,1H, Fuc H-3), 3.96 (s, 1H, Gal H-4), 4.03-4.08 (m, 1H, Lac H-2), 4.71(d, J=8.1 Hz, 1H, Gal H-1), 4.93-4.97 (m, 1H, Fuc H-1), 4.99 (dd, J=6.1,12.6 Hz, 1H, Fuc H-5), 5.44 (t, J=8.8 Hz, 1H, Gal H-2), 7.46-7.52 (m,2H, C₆H₅), 7.58-7.64 (m, 1H, C₆H₅), 8.04-8.11 (m, 2H, C₆H₅); ¹³C-NMR(125 MHz, MeOD): δ 16.70 (Fuc C-6), 19.20 (Me), 26.53 (Cy), 26.72 (Cy),27.28 (Cy), 33.08 (Cy), 34.17 (Cy), 35.05, 35.08 (2C, Cy, C-2), 37.10(C-6), 39.20 (C-5), 41.38 (C-1), 42.78 (Lac C—), 62.68 (Gal C-6), 67.70,67.75 (2C, Gal C-4, Fuc C-5), 70.32 (Fuc C-2), 71.44 (Fuc C-3), 73.01(Gal C-2), 73.97 (Fuc C-4), 75.89 (Gal C-5), 77.91 (Lac C-2), 79.95(C-3), 83.16 (C-4), 83.67 (Gal C-3), 100.49 (Fuc C-1), 100.61 (Gal C-1),129.68 (C₆H₅), 130.92 (C₆H₅), 131.53 (C₆H₅), 134.35 (C₆H₅), 166.94(O(C═O)Ph), 177.88 (cHex COOH), 178.89 (Lac COOH); elemental analysiscalcd (%) for C₃₆H₅₂O₁₆.H₂O (758.80): C, 56.98; H, 7.14. found: C,57.09/57.08; H, 6.93/6.97. [a]_(D) ²⁰=−67.2 (c=0.40, MeOH); IR (KBr):ν=3433 (vs, OH), 2927 (s), 2854 (m) 1720 (vs, C═O), 1638 (w), 1450 (m),1339 (m), 1316 (m), 1272 (vs), 1207 (m), 1167 (m), 1111 (vs), 1079 (vs),999 (m), 967 (w), 846 (w), 806 (vw), 771 (vw) 712 (s), 677 (w), 628 (w),559 (w) cm¹.

General Procedure I for the Synthesis of Intermediates 16-19

The carboxyl group attached to the cyclohexanediole ring (cHex COOH) in15 can selectively be converted into an active ester using HBTU/HOBt inpresence of the lactic acid carboxyl group (Lac COOH). This finding wasexploited for the attachment of the alkyne linker to thecyclohexanediole ring as described below.

Dicarboxylic acid 15 (1 equiv.) and HOBt (3 equiv.) were dissolved inanhydrous DMF (approx. 0.025 M) under argon. HBTU (1.1-1.2 equiv.) wasadded and the solution was stirred at r.t. for 5 min. An excess ofalkyne amine (approx. 10% v/v) was added, and stirring was continueduntil no further consumption of starting material was observed in MS(after 1.5 to 3 h). The solvent was removed in vacuo, and the residuewas purified by preparative HPLC-MS (H₂O/MeCN+0.2% HCOOH), and thedesired monoamide was isolated based on its retention time(t_(R(cHex amide))>t_(R(Lac amide))). The identity of the amide wasverified by 2D HMBC spectra (coupling of linker and cyclohexyl protonsto the amide C═O).

General Procedure II for the Synthesis of Products 20-23

To an approx. 0.03 mm solution of TMS-protected alkyne amide (1 equiv.)in anhydrous THF, TBAF (2 equiv.; 1 M solution in THF) was added at r.t.under argon. The solution was stirred for 1 to 2 h, and the solvent wasremoved in vacuo. The residue was purified by RP-18 chromatography(water/MeOH) or preparative HPLC-MS (H₂O/MeCN+0.2% HCOOH).

Synthesis of Intermediate 16 (1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-[3-(trimethylsilyl)prop-2-yn-1-yl]cyclohexanecarboxamide

Following general procedure I, dicarboxylic acid 15 (100 mg, 0.135mmol), HOBt (55.2 mg, 0.408 mmol), HBTU (53.6 mg, 0.141 mmol) and alkyneamine 11 (500 μL) were stirred at r.t. for 3 h to afford the desiredmonoamide 16 as a white solid (41 mg, 35%). The starting material (1S)was partially recovered (28 mg, 28%).

¹H-NMR (500 MHz, MeOD): δ 0.15 (s, 9H, TMS-H), 0.52-0.75 (m, 4H, Cy),0.91 (d, J=12.6 Hz, 1H, Cy), 1.10 (d, J=6.3 Hz, 3H, Me), 1.14-1.39 (m,10H, H-2_(a), H-6_(a), 5Cy, Fuc H-6), 1.39-1.47 (m, 1H, Lac H-3_(a)),1.47-1.55 (m, 1H, Lac H-3_(b)), 1.55-1.71 (m, 3H, Cy, H-6_(b), H-5),2.12-2.18 (m, 1H, H-2_(b)), 2.27 (s, 1H, H-1), 3.14 (t, J=9.5 Hz, 1H,H-4), 3.57 (t, J=5.7 Hz, 1H, Gal H-5), 3.62-3.81 (m, 6H, Gal H-3, H-3,Fuc H-2, Fuc H-4, Gal H-6), 3.83-3.90 (m, 2H, Fuc H-3, H-1′_(a)),3.92-3.99 (m, 2H, Gal H-4, H-1′_(b)), 4.04 (dd, J=2.7, 9.5 Hz, 1H, LacH-2), 4.69 (d, J=8.0 Hz, 1H, Gal H-1), 4.94-5.00 (m, 2H, Fuc H-1, FucH-5), 5.43 (t, J=8.8 Hz, 1H, Gal H-2), 7.48-7.54 (m, 2H, C₆H₅),7.59-7.64 (m, 1H, C₆H₅), 8.03-8.08 (m, 2H, C₆H₅). ¹³C-NMR (125 MHz,MeOD): δ −0.07 (Me₃Si), 16.69 (Fuc C-6), 19.22 (Me), 26.55 (Cy), 26.68(Cy), 27.24 (Cy), 30.37 (C-1′), 33.12 (Cy), 34.17 (Cy), 35.02 (Cy),35.31 (C-2), 37.25 (C-6), 39.25 (C-5), 42.73 (Lac C-3), 42.87 (C-1),62.71 (Gal C-6), 67.68, 67.77 (2C, Fuc C-5, Gal C-4), 70.30 (Fuc C-2),71.43 (Fuc C-3), 72.99 (Gal C-2), 73.94 (Fuc C-4), 75.94 (Gal 5-C),77.99 (Lac C-2), 79.89 (C-3), 83.02 (C-4), 83.68 (Gal C-3), 87.81(C-3′), 100.41 (Fuc C-1), 100.54 (Gal C-1), 103.04 (C-2′), 129.74(C₆H₅), 130.87 (C₆H₅), 131.53 (C₆H₅), 134.34 (C₆H₅), 166.84 (O(C═O)Ph),176.41 (CONH), 178.81 (COOH); MS: m/z calcd. for C₄₂H₆₃NO₁₅Si [M−H]⁻:848.39. found: 848.54.

Synthesis of Intermediate 17(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-[4-(trimethylsilyl)but-3-yn-1-yl]cyclohexanecarboxamide

Following general procedure I, dicarboxylic acid 15 (31.7 mg, 0.0428mmol), HOBt (17.7 mg, 0.135 mmol), HBTU (19.1 mg, 0.0504 mmol) andalkyne amine 12 (200 μL) were stirred at r.t. for 1.5 h to afford thedesired monoamide 17 as a white solid (30 mg, 70%). The startingmaterial (1S) was partially recovered (6.5 mg, 18%).

¹H NMR (500 MHz, MeOD): δ 0.15 (s, 9H, TMS-H), 0.52-0.73 (m, 4H, Cy),0.91 (d, J=12.1 Hz, 1H, Cy), 1.11 (d, J=6.4 Hz, 3H, Me), 1.14-1.38 (m,10H, H-2_(a), H-6_(a), 5Cy, Fuc H-6), 1.38-1.46 (m, 1H, Lac H-3_(a)),1.47-1.71 (m, 4H, Lac H-3_(b), Cy, H-6_(b), H-5), 2.09-2.16 (m, 1H,H-2_(b)), 2.23-2.30 (m, 1H, H-1), 2.32-2.38 (m, 2H, H-2′), 3.11-3.20 (m,2H, H-4, H-1′_(a)), 3.20-3.28 (m, 1H, H-1′_(b)), 3.55 (t, J=5.8 Hz, 1H,Gal H-5), 3.63 (dd, J=2.8, 9.7 Hz, 1H, Gal H-3), 3.66-3.81 (m, 5H, H-3,Fuc H-2, Fuc H-4, Gal H-6), 3.86 (dd, J=3.2, 10.3 Hz, 1H, Fuc H-3), 3.97(d, J=2.1 Hz, 1H, Gal H-4), 4.05 (dd, J=2.6, 9.7 Hz, 1H, Lac H-2), 4.67(d, J=8.1 Hz, 1H, Gal H-1), 4.97-5.02 (m, 2H, Fuc H-1, Fuc H-5), 5.44(dd, J=8.5, 9.3 Hz, 1H, Gal H-2), 7.48-7.53 (m, 2H, C₆H₅), 7.60-7.65 (m,1H, C₆H₅), 7.94 (t, J=5.8 Hz, 1H, CONH), 8.04-8.09 (m, 2H, C₆H₅). ¹³CNMR (125 MHz, MeOD): δ 0.30 (Me₃Si), 16.70 (Fuc C-6), 19.26 (Me), 21.02(C-2′), 26.53 (Cy), 26.68 (Cy), 27.23 (Cy), 33.07 (Cy), 34.14 (Cy),35.02 (Cy), 35.51 (C-2), 37.33 (C-6), 39.24 (C-5), 39.29 (C-1′), 42.72(Lac C-3), 43.11 (C-1), 62.69 (Gal C-6), 67.66, 67.73 (2C, Gal C-4, FucC-5), 70.26 (Fuc C-2), 71.38 (Fuc C-3), 72.94 (Gal C-2), 73.93 (FucC-4), 75.94 (Gal C-5), 77.93 (Lac C-2), 79.86 (C-3), 82.96 (C-4), 83.62(Gal C-3), 86.23 (C-4′), 100.41 (Fuc C-1), 100.56 (Gal C-1), 105.35(C-3′), 129.73 (C₆H₅), 130.87 (C₆H₅), 131.48 (C₆H₅), 134.37 (C₆H₅),166.78 (O(C═O)Ph), 177.06 (CONH), 178.71 (COOH); MS: m/z: calcd. forC₄₃H₆₅NO₁₅Si [M−H]⁻: 862.41. found: 862.38.

Synthesis of Intermediate 18 (1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-[5-(trimethylsilyl)pent-4-yn-1-yl]cyclohexanecarboxamide

Following general procedure I, dicarboxylic acid 15 (120 mg, 0.159mmol), HOBt (67 mg, 0.496 mmol), HBTU (74 mg, 0.195 mmol) and alkyneamine 13 (520 μL) were stirred at r.t. for 2 h to afford the desiredmonoamide 18 as a white solid (57 mg, 40%). The starting material (1S)was partially recovered (23 mg, 19%).

¹H-NMR (500 MHz, MeOD): δ 0.13 (s, 9H, TMS-H), 0.51-0.62 (m, 1H, Cy),0.62-0.73 (m, 3H, Cy), 0.85-0.96 (m, 1H, Cy), 1.11 (d, J=6.5 Hz, 3H,Me), 1.15-1.38 (m, 10H, H-2_(a), H-6_(a), 5Cy, Fuc H-6), 1.37-1.44 (m,1H, Lac H-3_(a)), 1.47-1.55 (m, 1H, Lac H-3_(b)), 1.55-1.69 (m, 6H, Cy,H-6_(b), H-5, H-2′), 2.12 (d, J=11.1 Hz, 1H, H-2), 2.22 (t, J=7.2 Hz,3H, H-3′, H-1), 3.10-3.24 (m, 3H, H-1′, H-4), 3.56 (t, J=5.7 Hz, 1H, GalH-5), 3.62 (dd, J=2.9, 9.7 Hz, 1H, Gal H-3), 3.65-3.81 (m, 5H, H-3, FucH-2, Fuc H-4, Gal H-6), 3.87 (dd, J=3.3, 10.3 Hz, 1H, Fuc H-3), 3.96 (d,J=7.1 Hz, 2H, Lac H-2, Gal H-4), 4.68 (d, J=8.1 Hz, 1H, Gal H-1),4.95-5.02 (m, 2H, Fuc H-1, Fuc H-5), 5.43 (dd, J=8.3, 9.4 Hz, 1H, GalH-2), 7.48-7.53 (m, 2H, C₆H₅), 7.59-7.64 (m, 1H, C₆H₅), 7.77 (t, J=5.6Hz, 1H, CONH), 8.04-8.08 (m, 2H, C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 0.23(TMS-C), 16.72 (Fuc C-6), 18.03 (C-2′), 19.22 (Me), 26.56 (Cy), 26.70(Cy), 27.26 (Cy), 29.64 (C-3′), 33.09 (Cy), 34.15 (Cy), 35.06 (Cy), v(C-2), 37.35 (C-6), 39.33 (C-5), 39.43 (C-1′), 42.76 (Lac C-3), 43.34(C-1), 62.70 (Gal C-6), 67.69, 67.72 (Gal H-4, Fuc H-5), 70.28 (FucC-2), 71.40 (Fuc C-3), 72.96 (Gal C-2), 73.97 (Fuc C-4), 75.93 (Gal5-C), 77.77 (Lac C-2), 79.90 (C-3), 83.01 (C-4), 83.67 (Gal C-3), 85.42(C-5′), 100.49 (Fuc C-1), 100.56 (Gal C-1), 107.62 (C-4′), 129.79(C₆H₅), 130.87 (C₆H₅), 131.56 (C₆H₅), 134.41 (C₆H₅), 166.77 (O(C═O)Ph),177.12 (CONH), 178.75 (COOH); MS: calcd for C₄₄H₆₇NO₁₅Si [M−H]⁻: 876.42.found: 862.63. [a]_(D) ²⁰=−61.5 (c=0.79, MeOH); IR (KBr): ν=3430 (vs,OH), 2927 (s), 2854 (m), 2175 (vw), 1726 (s, C═O), 1650 (m, C═O), 1542(vw), 1535 (vw), 1450 (w), 1369 (w), 1338 (w), 1315 (w), 1271 (s), 1250(m), 1166 (w), 1140 (vs), 1079 (vs) 1036 (s), 1001 (w), 969 (vw), 844(m, Me₃Si), 760 (vw), 709 (w) cm⁻¹.

Synthesis of Intermediate 19(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-[6-(trimethylsilyl)hex-5-yn-1-yl]cyclohexanecarboxamide

Following general procedure I, dicarboxylic acid 15 (47 mg, 0.0634mmol), HOBt (26 mg, 0.192 mmol), HBTU (29 mg, 0.0765 mmol) and alkyneamine 14 (200 μL) were stirred at r.t. for 2 h to afford the desiredmonoamide 19 as a white solid (25 mg, 44%). The starting material (1S)was partially recovered (5.5 mg, 12%).

¹H-NMR (500 MHz, MeOD): δ 0.12 (s, 9H, TMS-H), 0.51-0.75 (m, 4H, Cy),0.86-0.96 (m, 1H, Cy), 1.11 (d, J=6.4 Hz, 3H, Me), 1.15-1.28 (m, 3H,H-6_(a), H-2_(a), Cy), 1.27-1.38 (m, 7H, Fuc H-6, Cy), 1.38-1.45 (m, 1H,Lac H3_(a)), 1.45-1.62 (m, 7H, Lac H3_(b), H-3′, H-2′, H-6_(b), Cy),1.62-1.71 (m, 1H, H-5), 2.08-2.14 (m, 1H, H-2_(b)), 2.21-2.28 (m, 3H,H-4′, H-1), 3.08-3.12 (m, 2H, H-1′), 3.15 (t, J=9.6 Hz, 1H, H-4), 3.56(t, J=5.9 Hz, 1H, Gal H-5), 3.65 (dd, J=2.7, 9.8 Hz, 1H, Gal H-3),3.67-3.81 (m, 5H, H-3, Fuc H-2, Gal H-6), 3.86 (dd, J=3.2, 10.3 Hz, 1H,Fuc H-3), 3.96 (d, J=2.3 Hz, 1H, Gal H-4), 4.06 (dd, J=2.7, 9.7 Hz, 1H,Lac H-2), 4.69 (d, J=8.0 Hz, 1H, Gal H-1), 4.95-5.01 (m, 2H, Fuc H-1,Fuc H-5), 5.43 (t, J=8.9 Hz, 1H, Gal H-2), 7.48-7.53 (m, 2H, C₆H₅),7.60-7.65 (m, 1H, C₆H₅), 7.77 (t, J=5.6 Hz, 1H, CONH), 8.05-8.08 (m, 2H,C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 0.28 (TMS-C), 16.71 (Fuc C-6), 19.25(Me), 20.07 (C-4′), 26.53 (Cy), 26.68 (Cy), 26.96 (C3′), 27.24 (Cy),29.54 (C-2′), 33.11 (Cy), 34.15 (Cy), 35.02 (Cy), 35.36 (C-2), 37.51(C-6), 39.33 (C-5), 39.70 (C-1′), 42.72 (Lac C-3), 43.31 (C-1), 62.71(Gal C-6), 67.69 (Gal C-4), 67.74 (Fuc C-5), 70.27 (Fuc C-2), 71.39 (FucC-3), 72.99 (Gal C-2), 73.95 (Fuc C-4), 75.92 (Gal C-5), 77.85 (LacC-2), 79.84 (C-3), 83.00 (C-4), 83.63 (Gal C-3), 85.22 (C-6′), 100.44(Fuc C-1), 100.52 (Gal C-1), 108.17 (C-5′), 129.76 (C₆H₅), 130.85(C₆H₅), 131.53 (C₆H₅), 134.40 (C₆H₅), 166.78 (O(C═O)Ph), 176.96 (CONH),178.72 (COOH).

Synthesis of Product 20(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(prop-2-yn-1-yl)cyclohexanecarboxamide

Following general procedure II, 16 (49 mg, 0.0576 mmol) was reacted withTBAF (115 μL, 0.115 mmol) in 2 mL anhydrous THF. After purification withpreparative HPLC-MS, 36 mg (80%) of 20 was obtained as a white solid.

¹H-NMR (500 MHz, MeOD): δ 0.52-0.75 (m, 4H, Cy), 0.85-0.97 (m, 1H, Cy),1.11 (d, J=6.4 Hz, 3H, Me), 1.14-1.27 (4 Cy, H-6_(a), H-2_(a)),1.27-1.38 (m, 4H, Cy), 1.34 (d, J=6.5 Hz, 3H, Fuc H-6a), 1.38-1.46 (m,1H, Lac H-3_(a)), 1.51 (ddd, J=3.9, 10.1, 13.7 Hz, 1H, Lac H-3_(b)),1.54-1.71 (m, 3H, H-6_(b), Cy, H-5), 2.11-2.18 (m, 1H, H-2_(b)),2.22-2.31 (m, 1H, H-1), 2.59 (t, J=2.3 Hz, 1H, H-3′), 3.14 (t, J=9.5 Hz,1H, H-4), 3.57 (t, J=5.9 Hz, Gal H-5), 3.65 (dd, J=2.94, 9.77 Hz, 1H,Gal H-3), 3.67-3.92 (m, 8H, H-3, Fuc H-4, 2 Gal H-6, Fuc H-3, 2 H-1′),3.96 (d, J=2.2 Hz, 1H, Gal H-4), 4.05 (dd, J=2.7, 9.6 Hz, 1H, Lac H-2),4.69 (d, J=8.0 Hz, 1H, Gal H-1), 4.96-5.00 (m, 2H, Fuc H-1, Fuc H-5),5.42-5.45 (m, 1H, Gal H-2), 7.48-7.51 (m, 2H, C₆H₅), 7.60-7.63 (m, 1H,C₆H₅), 8.05-8.06 (m, 2H, C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 16.70 (FucC-6), 19.22 (Me), 26.51 (Cy), 26.68 (Cy), 27.25 (Cy), 29.39 (C-1′),33.12 (Cy), 34.17 (Cy), 35.02 (Cy), 35.25 (C-2), 37.25 (C-6), 39.26(C-5), 42.72 (Lac C-3), 42.93 (C-1), 62.72 (Gal C-6), 67.69, 67.78 (2C,Fuc C-5, Gal C-4), 70.31 (Fuc C-2), 71.42 (Fuc C-3), 72.17 (C-3′), 72.98(Gal C-2), 73.95 (Fuc C-4), 75.94 (Gal 5-C), 77.96 (Lac C-2), 79.86(C-3), 80.67 (C-2′), 83.00 (C-4), 83.65 (Gal C-3), 100.42 (Fuc C-1),100.55 (Gal C-1), 129.74 ( ), 130.86 ( ), 131.51 ( ), 134.38 (6C, C₆H₅),166.85 (O(C═O)Ph), 176.52 (CONH), 178.75 (COOH); [a]_(D) ²⁰=−67.5(c=1.6, MeOH); IR (KBr): ν=3436 (vs, OH), 3308 (s, C≡C), 2927 (s), 2851(m), 1727 (s, C═O), 1651 (m, C═O), 1534 (w), 1450 (m), 1341 (m), 1314(m), 1270 (s), 1160 (m), 1111 (vs), 1079 (vs), 1030 (s), 966 (w), 773(w), 712 (m), 674 (w), 631 (w) cm⁻¹.

Synthesis of Product 21(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-N-(but-3-yn-1-O-4-[(α-L-fucopyranosyl)oxy]-5-methyl-cyclohexanecarboxamide

Following general procedure II, 17 (80 mg, 0.0925 mmol) was reacted withTBAF (185 μL, 0.185 mmol) in 3.5 mL anhydrous THF. After purificationwith RP-18 chromatography, 68 mg (93%) of 21 was obtained as a whitesolid.

¹H-NMR (500 MHz, MeOD): δ 0.53-0.76 (m, 4H, Cy), 0.86-0.97 (m, 1H, Cy),1.11 (d, J=6.3 Hz, 3H, Me), 1.14-1.27 (m, 3H, Cy, H-6_(a), H-2_(a)),1.27-1.38 (m, 4H, Cy), 1.34 (d, J=6.5 Hz, 3H, Fuc H-6), 1.38-1.46 (m,1H, Lac H-3_(a)), 1.46-1.54 (m, 1H, Lac H-3_(b)), 1.54-1.73 (m, 3H,H-6_(b), Cy, H-5), 2.13 (d, J=11.8 Hz, 1H, H-2_(b)), 2.21-2.32 (m, 4H,H-1, 2 H-2′, H-4′), 3.15 (t, J=9.5 Hz, 1H, H-4), 3.18-3.27 (m, 2H,H-1′), 3.54-3.59 (m, 1H, Gal H-5), 3.63-3.82 (m, 6H, Gal H-3, H-3, FucH-2, Fuc H-4, 2 Gal H-6), 3.87 (dd, J=3.1, 10.3 Hz, 1H, Fuc H-3), 3.96(d, J=1.8 Hz, 1H, Gal H-4), 4.05 (dd, J=2.7, 9.6 Hz, 1H, Lac H-1), 4.68(d, J=8.0 Hz, 1H, Gal H-1), 4.94-5.92 (m, 2H, Fuc H-1, Fuc H-5),5.41-5.45 (m, 1H, Gal H-2), 7.48-7.51, 7.61-7.64, (2 m, 3H, C₆H₅),8.06-8.07 (m, 2H, C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 16.69 (Fuc C-6),19.25 (Me), 19.77 (C2′), 26.50 (Cy), 26.67 (Cy), 27.24 (Cy), 33.13 (Cy),34.16 (Cy), 35.00 (Cy), 35.44 (C-2), 37.36 (C-6), 39.27 (C-5), 39.33(C-1′), 42.71 (Lac C-3), 43.18 (C-1), 62.72 (Gal C-6), 67.68, 67.77 (2C,Fuc C-5, Gal C-4), 70.31 (Fuc C-2), 70.78 (C-4′), 71.41 (Fuc C-3), 72.99(Gal C-2), 73.94 (Fuc C-4, 75.93 (Gal C-5), 77.96 (Lac C-2), 79.87(C-3), 82.22 (C3′), 82.98 (C-4), 83.61 (Gal C-3), 100.40 (Fuc C-1),100.56 (Gal C-1), 129.71, 130.86, 131.52, 134.36 (6C, C₆H₅), 166.79(O(C═O)Ph), 177.12 (CONH), 178.74 (COOH); [a]_(D) ²⁰=−72.8 (c=1.15,MeOH); IR (KBr): ν=3435 (s), 3311 (s, C≡C), 2927 (s), 2853 (m), 2115(vw), 1727 (s, C═O), 1648 (m, C═O), 1544 (w), 1450 (m), 1364 (m), 1340(m), 1315 (m), 1270 (s), 1221 (m), 1166 (m), 1111 (vs), 1073 (vs), 1031(s), 999 (m), 966 (m), 901 (vw), 867 (vw), 806 (vw), 770 (w), 712 (m),675 (w), 632 (w) cm⁻¹.

Synthesis of Product 22(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(pent-4-yn-1-yl)cyclohexanecarboxamide

Following general procedure II, 18 (77 mg, 0.0877 mmol) was reacted withTBAF (175 μL, 0.175 mmol) in 3.5 mL anhydrous THF. After purificationwith RP-18 chromatography, 69 mg (98%) of 22 was obtained as a whitesolid.

¹H-NMR (500 MHz, MeOD): δ 0.50-0.61 (m, 1H, Cy), 0.61-0.75 (m, 3H, Cy),0.86-0.97 (m, 1H, Cy), 1.11 (d, J=6.5 Hz, 3H, Me), 1.15-1.26 (m, 3H, Cy,H-6_(a), H-2_(a)), 1.27-1.38 (m, 4H, Cy), 1.34 (d, J=6.5 Hz, 3H, FucH-6) 1.39-1.49 (m, 1H, Lac H-3_(b)), 1.51 (ddd, J=3.8, 10.1, 13.7 Hz,1H, Lac H-3_(a)), 1.55-1.61 (m, 2H, Cy, H-6_(b)), 1.61-1.71 (m, 3H,H-2′, H-5), 2.08-2.15 (m, 1H, H-2), 2.18 (td, J=2.6, 7.2 Hz, 2H, H-3′),2.20-2.28 (m, 1H, H-1), 2.29 (t, J=2.6 Hz, 1H, H-5′), 3.14 (t, J=7.6 Hz,1H, H-4), 3.14-3.24 (m, 2H, H-1′), 3.54-3.59 (m, 1H, Gal H-5), 3.64-3.80(m, 6H, Gal H-3, H-3, Fuc H-2, Fuc H-4, 2 Gal H-6), 3.86 (dd, J=3.2,10.3 Hz, 1H, Fuc H-3), 3.96 (d, J=2.3 Hz, 1H, Gal H-4), 4.04 (dd, J=2.6,9.8 Hz, 1H, Lac H-2), 4.69 (d, J=8.0 Hz, 1H, Gal H-1), 4.96 (d, J=4.0Hz, 1H, Fuc H-1), 4.98-5.01 (m, 1H, Fuc H-5), 5.43 (dd, J=8.6, 9.2 Hz,1H, Gal H-2), 7.49-7.52, 7.61-7.64 (2 m, 3H, C₆H₅), 7.80 (t, J=5.6 Hz,1H, CONH), 8.05-8.07 (m, 2H, C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 16.66,16.71 (2C, C-3′, Fuc C-6), 19.23 (Me), 26.53, 26.70 (2C, Cy), 27.27(Cy), 29.48 (C-2′), 33.11, 34.17 (2C, Cy), 35.06, 35.47 (2C, Cy, C-2)),37.39 (C-6), 39.31, 39.39 (2C, C-1′, C-5), 42.79 (Lac C-3), 43.31 (C-1),62.75 (Gal C-6), 67.70, 67.74 (2C, Gal C-4, Fuc C-5), 70.10, 70.29 (2C,Fuc C-2, C-5′), 71.40 (Fuc C-3), 72.99 (Gal C-2), 73.96 (Fuc C-4), 75.95(Gal C-5), 78.01 (Lac C-2), 79.89 (C-3), 83.00 (C-4), 83.65 (Gal C-3),84.23 (C-4′), 100.46 (Fuc C-1), 100.57 (Gal C-1), 129.76, 130.85,131.54, 134.42 (6C, C₆H₅), 166.8 (O(C═O)Ph), 177.10 (CONH), 179.01(COOH); [a]_(D) ²°=−66.4 (c=2.07, MeOH); IR (KBr): ν=3443 (vs, OH), 2928(s, C≡C), 2115 (vw), 1726 (s, C═O), 1648 (m, C═O), 1544 (w), 1450 (m),1270 (s), 1166 (m), 1111 (s), 1078 (vs), 1031 (s), 966 (s), 712 (m)cm⁻¹.

Synthesis of Product 23(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-N-(hex-5-yn-1-yl)-5-methyl-cyclohexanecarboxamide

Following general procedure II, 19 (25 mg, 0.0280 mmol) was reacted withTBAF (84 μL, 0.0840 mmol) in 1 mL anhydrous THF. After purification withpreparative HPLC-MS, 20 mg (87%) of 23 was obtained as a white solid.

¹H-NMR (500 MHz, MeOD): δ 0.51-0.75 (m, 4H, Cy), 0.86-0.97 (m, 1H, Cy),1.11 (d, J=6.4 Hz, 3H, Me), 1.16-1.27 (m, 3H, H-6_(a), H-2_(a), Cy),1.27-1.38 (m, 7H, Fuc H-6, Cy), 1.38-1.45 (m, 1H, Lac H3_(a)), 1.46-1.62(m, 7H, Lac H3_(b), H-3′, H-2′, H-6_(b), Cy), 1.63-1.71 (m, 1H, H-5),2.08-2.15 (m, 1H, H-2_(b)), 2.16-2.30 (m, 4H, H-4′, H-6′, H-1),3.05-3.13 (m, 2H, H-1′), 3.15 (t, J=9.5 Hz, 1H, H-4), 3.57 (t, J=5.8 Hz,1H, Gal H-5), 3.63-3.82 (m, 6H, Gal H-3, H-3, Fuc H-2, Fuc H-4, GalH-6), 3.87 (dd, J=2.9, 10.3 Hz, 1H, Fuc H-3), 3.94-3.98 (m, 1H, GalH-4), 4.02-4.07 (m, 1H, Lac H-2), 4.69 (d, J=8.0 Hz, 1H, Gal H-1),4.95-5.03 (m, 2H, Fuc H-1, Fuc H-5), 5.44 (t, J=8.9 Hz, 1H, Gal H-2),7.47-7.53 (m, 2H, C₆H₅), 7.60-7.65 (m, 1H, C₆H₅), 8.04-8.09 (m, 2H,C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 16.71 (Fuc C-6), 18.70 (C-4′), 19.25(Me), 26.52 (Cy), 26.68 (Cy), 26.90 (C3′), 27.25 (Cy), 29.46 (C-2′),33.12 (Cy), 34.16 (Cy), 35.04 (Cy), 35.36 (C-2), 37.50 (C-6), 39.33(C-5), 39.68 (C-1′), 42.77 (Lac C-3), 43.32 (C-1), 62.74 (Gal C-6),67.69 (Gal C-4), 67.75 (Fuc C-5), 69.85 (C-6′), 70.29 (Fuc C-2), 71.40(Fuc C-3), 72.98 (Gal C-2), 73.94 (Fuc C-4), 75.94 (Gal C-5), 78.02 (LacC-2), 79.83 (C-3), 83.00 (C-4), 83.64 (Gal C-3), 84.70 (C-5′), 100.42(Fuc C-1), 100.51 (Gal C-1), 129.73 (C₆H₅), 130.85 (C₆H₅), 131.54(C₆H₅), 134.38 (C₆H₅), 166.79 (O(C═O)Ph), 176.96 (CONH), 178.99 (COOH);IR (KBr): ν=3436 (s, OH), 2928 (s), 2868 (m), 1731 (s, C═O), 1653 (m),1550 (w), 1451 (m), 1350 (w), 1298 (m), 1270 (s), 1114 (vs), 1078 (vs),1034 (s), 996 (w), 966 (w), 712 (m) cm⁻¹.

Example 2 Synthesis of the Indole Derivatives (FIG. 4 and FIG. 5)Synthesis of Intermediate 24 1-(2-Bromoethyl)-5-nitro-1H-indole

5-nitro-1H-indole (1.00 g, 6.17 mmol) was added to powdered KOH (431 mg,7.68 mmol) dissolved in DMF (40 mL) at r.t. To this, 1,2-dibromoethane(1.55 mL, 18.5 mmol) was quickly added, and the solution was stirred for25 h. Water (100 mL) was added to the reaction mixture, and it wasextracted with Et₂O (3×100 mL). The organic phases were washed with 0.5M aq. HCl, satd. NaHCO₃, and brine (60 mL). The organic layer was driedover Na₂SO₄, and the solvent was removed under reduced pressure. Silicagel chromatography (EtOAc in petrol ether, gradient 0 to 85%) afforded24 (198 mg, 19%) as a yellow solid. Approximately 600 mg of the startingmaterial was recovered.

¹H-NMR (500 MHz, CDCl₃): δ 3.68 (t, J=6.6 Hz, 2H, H-1′), 4.59 (t, J=6.6Hz, 2H, H-2′), 6.71-6.74 (m, 1H, H-3), 7.29-7.33 (m, 1H, H-2), 7.36-7.40(m, 1H, H-7), 8.13-8.17 (m, 1H, H-6), 8.59-8.64 (m, 1H, H-4).

Synthesis of Intermediate 25 1-(3-Bromopropyl)-5-nitro-1H-indole

5-nitro-1H-indole (2.01 g, 12.3 mmol) was added to powdered KOH (0.692g, 12.3 mmol) dissolved in DMF (100 mL) at r.t., leading to a redsolution. To this, dibromopropane (3.77 mL, 37.0 mmol) was quicklyadded, accompanied by a color change to yellow. After 40 min, additional0.5 mL (4.90 mmol) of dibromopropane was added, whereupon someprecipitation occurred. After 17 h of stirring at r.t., water (200 mL)was added to the reaction mixture, and it was extracted with Et₂O (3×200mL). The organic phases were washed with 0.5 M aq. HCl, satd. NaHCO₃,and brine (100 mL). The organic layers were dried over Na₂SO₄, and thesolvent was removed under reduced pressure. Silica gel chromatography(EtOAc in petrol ether, gradient 0 to 85%) afforded 25 as a yellowcrystals (1.59 g, 46%). 0.565 g (28%) of the starting material wasrecovered.

¹H-NMR (500 MHz, CDCl₃): δ 2.35-2.41 (m, 2H, H-2′), 3.30-3.33 (m, 2H,H-3′), 4.40 (t, J=6.5 Hz, 2H, H-1′), 6.70 (m, 1H, H-3), 7.30-7.32 (m,1H, H-2), 7.40-7.43 (m, 1H, H-7), 8.10-8.13 (m, 1H, H-6), 8.57 (m, 1H,H-4); ¹³C-NMR (125 MHz, CDCl₃): δ 30.05 (C-3′), 32.70 (C-2′), 44.53(C-1′), 104.54 (C-3), 109.33 (C-7), 117.52 (C-6), 118.42 (C-4), 127.92(C-9), 131.22 (C-2), 138.84 (C-8), 141.79 (C-5); elemental analysiscalcd (%) for C₁₁H₁₁BrN₂O₂ (283.12): C, 53.87; H, 4.52; N, 28.56. found:C, 53.86/53.77; H, 4.71/4.65; N, 28.29/28.32. IR (KBr): ν=1606 (vw),1577 (vw), 1511 (s, NO₂), 1480 (m), 1459 (m), 1431 (vw), 1404 (w), 1331(vs, NO₂), 1296 (s), 1253 (m), 1226 (m), 1183 (w), 1157 (w), 1068 (m),975 (vw), 934 (vw), 902 (w), 816 (w), 768 (w), 749 (s) cm⁻¹.

Synthesis of Intermediate 26 1-(4-Bromobutyl)-5-nitro-1H-indole

To a solution of 5-nitro-1H-indole (2.00 g, 12.3 mmol) in anhydrous

EtOAc (20 mL) was added K₂CO₃ (3.39 g, 24.5 mmol) and TBAB (4.76 g, 14.8mmol). The suspension was stirred at 50° C. for 30 min followed by theaddition of 1,4-dibromobutane (1.74 mL, 14.8 mmol). Stirring wascontinued under reflux for 21 h. Upon completion of the reaction, water(20 mL) was added to the reaction mixture, the phases were separated,and the aqueous phase was further extracted with EtOAc (2×40 mL). Theorganic layers were washed with 0.5 M aq. HCl, satd. NaHCO₃, and brine(20 mL), subsequently dried over Na₂SO₄ and concentrated. The residuewas purified using silica gel chromatography (EtOAc in petrol ether) toafford bromide 26 as a yellow solid (1.67 g, 46%).

¹H-NMR (500 MHz, CDCl₃): δ 1.83-1.90 (m, 2H, H-2′), 2.02-2.09 (m, 2H,H-3′), 3.40 (t, J=6.4 Hz, 2H, H-4′), 4.22 (t, J=7.0 Hz, 2H, H-1′),6.68-6.71 (m, 1H, H-3), 7.24-7.25 (m, 1H, H-2), 7.34-7.38 (m, 1H, H-7),8.11-8.15 (m, 1H, H-6), 8.58-8.61 (m, 1H, H-4).

Synthesis of Product 27 1-(2-Azidoethyl)-5-nitro-1H-indole

NaN₃ (69.6 mg, 1.07 mmol) was added to a solution of bromide 24 (198 mg,0.736 mmol) in DMF (2 mL), and the resulting suspension was stirred at60° C. After 12 h, water (10 mL) was added, and the reaction mixture wasextracted with EtOAc (3×20 mL). The organic layers were washed withwater and brine (10 mL) and subsequently dried over Na₂SO₄. Silica gelchromatography (EtOAc in petrol ether) afforded azide 27 (90 mg, 52%) asa yellow solid.

¹H-NMR (500 MHz, DMSO-d6): δ 3.76 (t, J=5.6 Hz, 2H, H-2′), 4.48 (t,J=5.6 Hz, 2H, H-1′), 6.77-6.83 (m, 1H, H-3), 7.66-7.70 (m, 1H, H-2),7.74-7.80 (m, 1H, H-7), 8.02-8.08 (m, 1H, H-6)), 8.54-8.62 (m, 1H, H-4);¹³C-NMR (125 MHz, DMSO-d6): δ 45.32 (C-1′), 50.66 (C-2′), 104.18 (C-3),110.51 (C-7), 116.53 (C-6), 117.59 (C-4), 127.50 (C-9), 132.56 (C-2),138.95 (C-8), 140.86 (C-5).

Synthesis of Product 28 1-(3-Azidopropyl)-5-nitro-1H-indole

NaN₃ (434 mg, 6.68 mmol) was added to a solution of bromide 25 (1.34 g,4.73 mmol) in DMF (10 mL), and the resulting suspension was stirred at55° C. After 12 h, water (20 mL) was added, and the reaction mixture wasextracted with EtOAc (3×30 mL). The organic layers were washed withwater and brine (20 mL) and subsequently dried over Na₂SO₄. Silica gelchromatography (EtOAc in petrol ether) afforded azide 28 (950 mg, 82%)as a yellow solid.

¹H-NMR (500 MHz, DMSO-d6): δ 2.03 (p, J=6.8 Hz, 2H, H-2′), 3.32 (t,J=6.7 Hz, 2H, H-3′), 4.34 (t, J=7.0 Hz, 2H, H-1′), 6.76-6.79 (m, 1H,H-3), 7.65-7.69 (m, 1H, H-2), 7.69-7.75 (m, 1H, H-7), 8.01-8.07 (m, 1H,H-6), 8.58 (s, 1H, H-4); ¹³C-NMR (125 MHz, DMSO-d6): δ 28.97 (C-2′),43.30 (C-1′), 48.00 (C-3′), 103.91 (C-3), 110.32 (C-7), 116.46 (C-6),117.64 (C-4), 127.36 (C-9), 132.50 (C-2), 138.68 (C-5), 140.73 (C-5).

Synthesis of Product 29 1-(4-Azidobutyl)-5-nitro-1H-indole

NaN₃ (531 mg, 8.17 mmol) was added to a solution of bromide 26 (1.60 g,5.38 mmol) in DMF (5 mL), and the resulting suspension was stirred at55° C. After 14 h, water (10 mL) was added, and the reaction mixture wasextracted with EtOAc (3×20 mL). The organic layers were washed withwater and brine (10 mL) and subsequently dried over Na₂SO₄. Silica gelchromatography (EtOAc in petrol ether) afforded azide 29 (1.10 g, 79%)as a yellow solid.

¹H-NMR (500 MHz, DMSO-d6): δ 1.43-1.50 (m, 2H, H-3′), 1.78-1.85 (m, 2H,H-2′), 3.31-3.38 (m, 2H, H-4′), 4.30 (t, J=7.0 Hz, 2H, H-1′), 6.74-6.77(m, 1H, H-3), 7.66-7.68 (m, 1H, H-2), 7.71-7.75 (m, 1H, H-7), 8.00-8.05(m, 1H, H-6), 8.56-8.58 (m, 1H, H-4); ¹³C-NMR (125 MHz, DMSO-d6): δ25.63 (C-3′), 27.10 (C-2′), 45.44 (C-1′), 50.15 (C-4′), 103.70 (C-3),110.40 (C-7), 116.37 (C-6), 117.64 (C-4), 127.32 (C-9), 132.55 (C-2),138.67 (C-8), 140.66 (C-5); elemental analysis calcd (%) for C₁₂H₁₃N₅O₂(259.26): C, 55.59; H, 5.05; N, 27.01. found C, 55.66/55.64; H,5.20/5.21; N, 26.81/26.79.

Synthesis of Intermediate 30 (5-Nitro-1H-indol-1-yl)methanol

5-nitro-1H-indole (995 mg, 6.14 mmol) and K₂CO₃ (396 mg, 2.87 mmol) weredissolved in EtOH in a bomb tube. The solution was stirred at 60° C. for5 min, and 8 mL of freshly prepared ˜30% aq. methanal (preparation: 3.00g paraformaldehyde and 0.901 g of K₂CO₃ were suspended in 10 mL H₂O andstirred at 60° C. until complete dissolution of the paraformaldehyde)were added. Stirring was continued at 60° C. for another 20 min. Afterremoval of the solvent, the crude mixture was redissolved in EtOAc andadsorbed on silica gel. The adsorbed material was subjected to silicagel chromatography (EtOAc in petrol ether, gradient 30 to 70%) to affordalcohol 30 as a yellow solid (784 mg, 66%).

¹H-NMR (500 MHz, MeOD): δ 5.62 (s, 2H, H-1′), 6.67-6.72 (m, 1H, H-3),7.48-7.52 (m, 1H, H-2), 7.60-7.65 (m, 1H, H-7), 8.05-8.10 (m, 1H, H-6),8.50-8.54 (m, 1H, H-4); ¹³C-NMR (125 MHz, MeOD): δ 70.45 (C-1′), 105.21(C-3), 111.17 (C-7), 117.95 (C-6), 118.54 (C-4), 129.92 (C-9), 132.41(C-2), 140.12 (C-8), 143.16 (C-5); elemental analysis calcd (%) forC₉H₈N₂O₃ (192.17): C, 56.25; HH, 4.20; N, 14.58. found: C, 56.14/56.35;HH, 4.21/4.20; N, 14.60/14.72. IR (KBr): ν=3486 (vs, OH), 1609 (w), 1580(vw), 1512 (s, NO₂), 1466 (m), 1399 (w), 1331 (vs, NO₂), 1279 (s), 1206(w), 1147 (vw), 1100 (w), 1073 (w), 1040 (s), 988 (vw), 894 (vw), 814(vw), 763 (vw), 744 (s), 722 (vw) cm⁻¹.

Synthesis of Product 31 1-(Azidomethyl)-5-nitro-1H-indole

Alcohol 30 (770 mg, 4.01 mmol) was dissolved in anhydrous THF (13 mL)under argon. At −15° C. (ice-salt bath), MeSO₂Cl (0.312 mL, 4.01 mmol)and Et₃N (1.1 mL, 8.02 mmol) were added. After 10 min, the ice bath wasremoved and stirring was continued for 1 h, when 15-Crown-5 (0.397 mL,2.01 mmol) and NaN₃ (521 mg, 8.02 mmol) were added to the solution.After another 5 h, the solvent was removed. The crude mixture wasdissolved in EtOAc (75 mL) and H₂O (50 mL). The two phases wereseparated, and the organic phase was washed with brine (50 mL). Theaqueous phase was extracted with EtOAc (2×50 mL). The combined organiclayers were dried over Na₂SO₄, and the solvent was removed under reducedpressure. After silica gel chromatography (EtOAc in petrol ether,gradient 20 to 70%), the azide 31 was obtained as yellow crystals (297mg, 34%). Furthermore, 30 mg of 5-nitro-1H-indole and 42 mg (5%) of thestarting material were isolated.

¹H-NMR (500 MHz, CDCl₃): δ 5.51 (s, 2H, H-1′), 6.75-6.79 (m, 1H, H-3),7.33-7.35 (m, 1H, H-2), 7.47-7.50 (m, 1H, H-7), 8.18-8.21 (m, 1H, H-6),8.60-8.62 (m, 1H, H-4); ¹³C-NMR (125 MHz, CDCl₃): δ 61.56 (C-1′), 106.03(C-3), 109.68 (C-7), 118.43 (C-6), 118.50 (C-4), 128.73 (C-9), 130.71(C-2), 138.88 (C-8), 142.84 (C-5); elemental analysis calcd (%) forC₉H₇N₅O₂ (217.18): C, 49.77; H, 3.25; N, 32.25. found: C, 49.83/49.59;H, 3.28/3.28; N, 32.56/32.46. IR (KBr): ν=3101 (vw), 2924 (vw), 2118 (m,N₃), 2092 (m, N₃), 1612 (w), 1583 (vw), 1509 (s, NO₂), 1480 (m), 1451(m), 1401 (vw), 1336 (vs, NO₂/N₃), 1287 (vs, N₃), 1266 (m), 1231 (m),1208 (w), 1179 (m), 1147 (w), 1076 (w), 1065 (w), 1029 (vw), 934 (vw),906 (w), 876 (w), 821 (w), 745 (s) cm⁻¹.

Synthesis of Intermediate 32 5-Nitro-1H-indole-3-carbaldehyde

Intermediate 32 was synthesized according to the procedure described in[S. Shelke, B. Cutting, X. Jiang, H. Koliwer-Brandl, D. S. Strasser, S.Kelm, O, Schwardt, B. Ernst, Angew. Chem. Int. Ed. 2010, 49,5721-5725.].

¹H-NMR (500 MHz, DMSO-d6): δ 7.71-7.74 (m, 1H, H-7), 8.14-8.17 (m, 1H,H-6), 8.57 (s, 1H, H-2), 8.94-8.96 (m, 1H, H-4), 10.03 (s, 1H, H-1′);¹³C-NMR (125 MHz, CDCl₃): δ 113.29 (C-7), 117.07 (C-4), 118.83 (C-3),119.07 (C-6), 123.55 (C-9), 140.17 (C-2), 141.51 (C-5), 142.91 (C-2),185.59 (C-1′).

Synthesis of Intermediate 33 (5-Nitro-1H-indol-3-yl)methanol

Intermediate 33 was synthesized according to the procedure described in[S. Shelke, B. Cutting, X. Jiang, H. Koliwer-Brandi, D. S. Strasser, S.Kelm, O. Schwardt, B. Ernst, Angew. Chem. Int. Ed. 2010, 49,5721-5725.].

¹H-NMR (500 MHz, MeOD): δ 4.84 (s, 2H, H-1′), 7.43 (s, 1H, H-2),7.45-7.49 (m, 1H, H-7), 8.03-8.07 (m, 1H, H-6), 8.66-8.69 (m, 1H, H-4);¹³C-NMR (125 MHz, MeOD): δ 56.74 (C-1′), 112.46 (C-7), 117.20 (C-4),118.02 (C-6), 119.29 (C-3), 127.53 (C-9), 128.04 (C-2), 141.38 (C-8),142.56 (C-5); elemental analysis calcd (%) for C₉H₈N₂O₃ (192.17): C,56.25; H, 4.20; N, 14.58. found: C, 56.18/56.00; H, 4.41/4.45; N,14.65/14.60.

Synthesis of Product 34 3-(Azidomethyl)-5-nitro-1H-indole

To alcohol 33 (752 mg, 3.91 mmol) suspended in toluene (10 mL) underargon were added diphenyl phosphoryl azide (1.0 mL, 4.69 mmol) anddropwise at −10° C. DBU (0.7 mL, 4.69 mmol). The suspension was allowedto warm to r.t., and 5 mL THF were added leading to the formation of twophases. The reaction was continued under vigorous stirring for 18 h.After removal of the THF in vacuo, EtOAc (80 mL) was added to thereaction mixture, and the organic phases were washed with satd. aq.NaHCO₃ (2×40 mL), 0.5 M aq. HCl (40 mL), satd. aq. NaHCO₃ (40 mL) andbrine (40 mL). The aqueous layers were extracted with EtOAc (2×80 mL),and the organic phase was dried over Na₂SO₄. Silica gel chromatography(1. CH₂Cl₂/MeOH 10:1 in CH₂Cl₂, gradient 0 to 100%; 2. CH₂Cl₂) affordedonly impure product. Subsequent crystallization from CH₂Cl₂ affordedpure azide 34 as yellow crystals (658 mg, 78%). 140 mg (19%) of thestarting material was recovered.

¹H-NMR (500 MHz, DMSO-d6): δ 4.70 (s, 2H, H-1′), 7.56-7.62 (m, 1H, H-7),7.75 (s, 1H, H-2), 8.01-8.08 (m, 1H, H-6), 8.65 (s, 1H, H-4), 11.90 (s,1H, NH); ¹³C-NMR (125 MHz, DMSO-d6): δ 45.16 (C-1′), 111.57 (C-3),112.38 (C-7), 115.76 (C-4), 117.07 (C-6), 125.88 (C-9), 129.50 C-2),139.53 (C-8), 140.88 (C-5); elemental analysis calcd (%) for C₉H₇N₅O₂(217.18): C, 49.77; H, 3.25; N, 32.25. found: C, 49.61/49.73; H,3.31/3.27; N, 32.07/32.14. IR (KBr): ν=3257 (m, NH), 2117 (vs, N₃), 1625(w), 1580 (w), 1551 (m), 1515 (s, NO₂), 1472 (m), 1456 (m), 1436 (m),1376 (w), 1331 (vs, N₃/NO₂), 1261 (m), 1224 (m), 1196 (m), 1128 (w),1113 (m), 1102 (m), 1045 (w), 976 (w), 939 (vw), 920 (vw), 894 (m), 863(w), 836 (m), 816 (m), 782 (w), 769 (w), 751 (m), 735 (m), 686 (m), 631(m), 613 (w), 563 (m), 559 (m) cm⁻¹.

Synthesis of Intermediate 35 tert-ButylN-[1-(3-bromopropyl)-1H-indol-5-yl]carbamate

Nitroindole 25 (700 mg, 2.50 mmol) and PtO₂ (35 mg, 5% w/w) weresuspended in a solution of Boc₂O (1.76 g, 8.06 mmol) in EtOH (23 mL)under argon. The flask was flushed with H₂, and the solution was stirredunder H₂ (atm. press.) at r.t. for 45 min (color change from yellow togreen), when another 35 mg of PtO₂ was added. After stirring foradditional 30 min, the now colorless mixture was filtrated (PTFEmembrane filter) and the solvent was removed in vacuo. The residue wasdissolved in EtOAc (100 mL) and washed with 0.5 M aq. HCl (50 mL), satd.aq. NaHCO₃ (50 mL), and brine (50 mL). The organic layer was dried overNa₂SO₄, and the solvent was removed under reduced pressure. Afterpurification by silica gel chromatography (EtOAc in petrol ether,gradient 0 to 25%), 35 (637 mg, 72%) was obtained as a pale yellow oil,which eventually crystallized and turned pink after 24 h at −18° C.

¹H-NMR (500 MHz, MeOD): δ 1.50 (s, 9H, ^(t)Bu-H), 2.28 (p, J=6.4 Hz, 2H,H-2′), 3.28 (m, 2H, H-3′), 4.27 (t, J=6.5 Hz, 2H, H-1′), 6.35 (m, 1H,H-3), 7.10 (m, 1H, H-6), 7.16 (m, 1H, H-2), 7.30 (m, 1H, H-7), 7.57 (s,1H, H-4); ¹³C-NMR (125 MHz, MeOD): δ 28.80 (^(t)Bu-CH₃), 31.16 (C-3′),34.31 (C-2′), 45.05 (C-1′), 80.44 (^(t)Bu-CCH₃), 102.08 (H-3), 110.37(C-7), 112.84 (C-6), 116.67 (C-4), 129.81 (C-2), 130.26 (C-9), 132.41(C-8), 134.36 (C-5), 156.27 (OCONH); elemental analysis calcd (%) forC₁₆H₂₁BrN₂O₂ (353.25): C, 54.40; H, 5.99; N, 7.93. found: C,54.29/54.35; H, 5.97/5.96; N, 7.86/7.76. IR (KBr): ν=2972 (w) 2932 (w),1695 (vs, C═O), 1625 (vw), 1584 (m), 1532 (s), 1509 (m), 1490 (s), 1447(m), 1441 (m), 1392 (m), 1367 (s), 1334 (m), 1312 (vw), 1295 (m), 1287(m), 1261 (m), 1232 (s), 1163 (vs), 1092 (vw), 1053 (m), 1027 (vw), 889(vw), 863 (vw), 825 (vw), 802 (vw), 756 (w), 726 (w), 623 (vw), 563 (vw)cm⁻¹.

Synthesis of Intermediate 36 tert-ButylN-[1-(3-azidopropyl)-1H-indol-5-yl]carbamate

To a solution of 35 (570 mg, 1.61 mmol) in anhydrous DMF (15 mL), NaN₃(528 mg, 8.12 mmol) was added at r.t. After 2 h, H₂O (50 mL) was addedto the reaction mixture, and the aqueous layers were extracted withEtOAc (100 mL). The organic layers were washed with satd. NaHCO₃, brine,and dried over Na₂SO₄. After removal of the solvent under reducedpressure, the crude product was purified by silica gel chromatography(EtOAc in petrol ether, gradient 0 to 25%) to afford 36 (458 mg, 90%) asslightly yellowish crystals.

¹H-NMR (500 MHz, MeOD): δ 1.52 (s, 9H, ^(t)Bu-H), 2.02 (p, J=6.6 Hz, 2H,H-2′), 3.22 (t, J=6.5 Hz, 2H, H-3′), 4.22 (t, J=6.7 Hz, 2H, H-1′),6.36-6.39 (m, 1H, H-3), 7.10-7.15 (m, 1H, H-6), 7.15-7.17 (m, 1H, H-2),7.28-7.32 (m, 1H, H-7), 7.59 (s, 1H, H-4). ¹³C-NMR (125 MHz, MeOD): δ28.80 (^(t)Bu-CH₃), 30.51 (C-2′), 44.02 (C-1′), 49.54 (C-3′), 80.40(^(t)Bu-CCH₃), 102.01 (H-3), 110.27 (C-7), 112.79 (C-6), 116.63 (C-4),129.72 (C-2), 130.20 (C-9), 132.32 (C-8), 134.38 (C-5), 156.24 (OCONH);elemental analysis calcd (%) for C₁₆H₂₁N₅O₂ (315.37): C, 60.94; H, 6.71;N, 22.21. found: 61.07/61.04; H, 6.69/6.70; N, 22.18/22.17. IR (KBr):ν=2976 (w), 2930 (w), 2873 (vw), 2099 (vs, N₃), 1697 (s, C═O), 1625(vw), 1584 (w), 1529 (m), 1510 (m), 1489 (s), 1450 (m), 1440 (m), 1392(w), 1366 (m), 1334 (w), 1286 (m), 1236 (m), 1159 (s), 1096 (vw), 1085(vw), 1051 (w), 1051 (m), 1026 (w) cm⁻¹.

Synthesis of Intermediate 37 1-(3-Azidopropyl)-1H-indol-5-amine

Carbamate 36 (408 mg, 1.29 mmol) was dissolved in CH₂Cl₂ (40 mL), andtrifluoroacetic acid (3 mL) was added. After stirring at r.t. for 30min, the solvent was removed in vacuo, and the mixture was co-evaporatedwith toluene. The residue was dissolved in EtOAc (50 mL), the organiclayer was washed with satd. aq. NaHCO₃ and brine (30 mL), and theaqueous layers were extracted with EtOAc (50 mL). The crude productobtained after drying the combined organic phase over Na₂SO₄ and removalof the solvent under reduced pressure was subjected to silica gelchromatography (MeOH in CH₂Cl₂ 10:1). The amine 37 (239 mg, 86%) wasobtained as a mixture of inseparable products (brownish oil).

¹H-NMR (500 MHz, MeOD): δ 1.52 (s, 9H, ^(t)Bu-H), 2.02 (p, J=6.6 Hz, 2H,H-2′), 3.22 (t, J=6.5 Hz, 2H, H-3′), 4.22 (t, J=6.7 Hz, 2H, H-1′), 6.37(m, 1H, H-3), 7.13 (m, 1H, H-6), 7.16 (m, 1H, H-2), 7.30 (m, 1H, H-7),7.59 (s, 1H, H-4).

Synthesis of Product 38 N-[1-(3-Azidopropyl)-1H-indol-5-yl]acetamide

To a solution of amine 37 (43 mg, 0.200 mmol) was added Et₃N (28 μL,0.200 mmol) and Ac₂O (18 μL, 0.200 mmol), and the solution was stirredfor 3 h. The solvent was removed in vacuo and the residue purified usingsilica gel chromatography (EtOAc in CH₂Cl₂). 38 (44 mg, 86%) wasobtained as a mixture of inseparable products (brownish oil).

¹H-NMR (500 MHz, CDCl₃): δ 1.92-1.96 (m, 2H, H-2′), 2.07 (s, 3H, Ac-Me),3.12 (t, J=6.3 Hz, 2H, H-3′), 4.09 (t, J=6.6 Hz, 2H, H-1′), 6.34-6.35(m, 1H, H-3), 6.97-6.99 (m, 1H, H-2), 7.14-7.16 (m, 2H, H-6, Ind H-7),7.70 (s, 1H, H-4); ¹³C-NMR (125 MHz, CDCl₃): δ 24.38 (Ac-Me), 29.35(C-2′), 43.14 (C-1′), 48.33 (C-3′), 101.72 (C-3), 109.33 (C-7), 113.41(C-4), 116.48 (C-6), 128.71, 128.72 (2C, C-2, C-9), 130.38 (C-8), 133.42(C-5), 168.85 (MeCONH). MS: m/z calcd for C₁₃H₁₅N₅O [M+Na]⁺: 280.12.found: 280.20.

Synthesis of Intermediate 39 2,5-Dioxopyrrolidin-1-yl2-[2-(2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}ethoxy)ethoxy]acetate

2-[2-(2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}ethoxy)ethoxy]aceticacid (185 mg, 0.480 mmol) and N-hydroxysuccinimide (110 mg, 0.956 mmol)were dissolved in anhydrous THF (1.5 mL). DIC was added dropwise to thesolution at r.t., leading to the precipitation of a white solid. After 2h, the reaction mixture was filtrated through a PTFE membrane filter(0.2 μm), and the solvent was removed. The crude product was redissolvedin Et₂O. The organic phase was washed with aq. NaHCO₃ and 0.5 M aq. HCl.The organic layer was dried over Na₂SO₄, and the solvent was removed.The crude product was dissolved in CH₂Cl₂, and the solution wasfiltrated. After removal of the solvent, the crude product 39 (210 mg)was used in the synthesis of 40 without further purification.

Synthesis of Product 409H-Fluoren-9-ylmethyl-N-{2-[2-({[1-(3-azidopropyl)-1H-indol-5-yl]carbamoyl}methoxy)ethoxy]ethyl}carbamate

Amine 37 (50 mg, 0.232 mmol) and succinimide ester 39 (120 mg, 0.249mmol) were dissolved in anhydrous DMF (1 mL) and stirred at r.t. for 15h. The solvent was removed in vacuo, and the residue was purified usingsilica gel chromatography (EtOAc in CH₂Cl₂, gradient 0 to 80%). 70 mg(52%) of 40 was obtained as mixture of inseparable products(green-yellow oil).

¹H-NMR (500 MHz, CDCl₃): δ 1.88-1.96 (m, 2H, H-2′), 3.10 (t, J=6.3 Hz,2H, H-3′), 3.32-3.38 (m, 2H, H-6″), 3.56 (t, J=5.0 Hz, 2H, H-5″), 3.64(d, J=4.2 Hz, 2H, H-4″), 3.73 (d, J=4.4 Hz, 2H, H-3″), 4.02-4.11 (m, 5H,H-9″, H-2″. H-1′), 4.25 (d, J=7.0 Hz, 2H, H-8″), 6.36-6.40 (m, 1H, IndH-3), 6.96-6.99 (m, 1H, Ind H-2), 7.15-7.25 (m, 4H, Ind H-6. Ind H-7,B), 7.27-7.34 (m, 2H, C), 7.42-7.48 (m, 2H, A), 7.64-7.71 (m, 2H, D),7.79-7.82 (m, J=1.4 Hz, 1H, Ind H-4), 8.56 (s, 1H, CONH); ¹³C-NMR (125MHz, CDCl₃): δ 29.43 (C-2′), 40.82 (C-6″), 43.23 (C-1′), 47.31 (C-9″),48.40 (C-3′), 66.83 (C-8″), 70.23, 70.43, 70.90, 71.28, 101.91 (C-3),109.60 (C-7), 113.11 (C-4), 115.99 (C-6), 120.11 (Fmoc-C), 125.22(Fmoc-C), 127.19 (Fmoc-C), 127.83 (Fmoc-C), 128.91 (Fmoc-C), 128.93(Fmoc-C), 129.62 (C-8), 133.63 (C-5), 141.41 (Fmoc-C), 144.06 (Fmoc-C),156.60 (OCONH), 167.97 (CH₂CONH); MS: m/z calcd for C₃₂H₃₄N₆O₅ [M+Na]⁺:605.25. found: 605.22.

Synthesis of Intermediate 41tert-Butyldimethyl[(4-(trimethylsilyl)but-3-yn-1-yl)oxy]silane

To a solution of 4-trimethylsilyl-3-butyne-1-ol (854 mg, 6.0 mmol) inDMF (4 mL) was added imidazole (816 mg, 12.0 mmol), followed bytert-butyldimethylsilyl chloride (1.05 g, 7.2 mmol). After stirring atr.t. overnight, the reaction mixture was poured into 10% aq. NaHCO₃ andextracted with hexane (3×10 mL). The combined organic phases were washedwith brine, dried over Na₂SO₄ and concentrated to give 41 (1.53 g,quant.) as a colorless oil.

¹H NMR (500 MHz, CDCl₃): δ 0.09 (s, 6H, 2 CH₃), 0.14 (s, 9H, Si(CH₃)₃),0.88 (s, 9H, C(CH₃)₃), 2.44 (t, J=7.0 Hz, 2H, CH₂), 3.72 (t, J=7.0 Hz,2H, CH₂).

Synthesis of Intermediate 42 tert-Butyl(hex-5-yn-1-yloxy)dimethylsilane

Hex-5-yn-1-ol (1.25 g, 12.7 mmol) was dissolved in anhydrous CH₂Cl₂under argon. At 0° C., Et₃N (2.6 mL, 19.1 mmol) andtert-butyldimethylsilyl chloride (2.29 g, 15.2 mmol) were added via asyringe. After 30 min of stirring, the solution was allowed to warm tor.t., which led to precipitation of a white solid. After another 4 h ofstirring, the reaction mixture was washed with 0.5 M aq. HCl, satd. aq.NaHCO₃, and brine (5 mL). The aqueous layers were washed with CH₂Cl₂(2×10 mL), and the combined organic phases were dried over Na₂SO₄,filtrated through a silica plug and again dried over Na₂SO₄. The solventwas removed in vacuo yielding 42 (2.31 g, 85%) as a colorless liquid,which was used without further purification.

¹H-NMR (500 MHz, MeOD): δ 0.04 (d, J=2.7 Hz, 6H, Si—CH₃), 0.89 (s, 9H,^(t)Bu-CH₃), 1.54-1.67 (m, 4H, H-3, H-2), 1.93 (t, J=2.6 Hz, 1H, H-6),2.21 (td, J=2.5, 6.8 Hz, 2H, H-4), 3.63 (t, J=6.0 Hz, 2H, H-1); ¹³C-NMR(125 MHz, MeOD): δ 5.24 (2C, Si—CH₃), 18.29 (C-4), 18.41 (^(t)Bu-C)25.04 (C-3), 26.03 (3C, ^(t)Bu-CH₃), 31.89 (C-2), 62.66 (C-1), 68.33(C-6), 84.59 (C-5).

Synthesis of Intermediate 43tert-Butyldimethyl[(6-(trimethylsilyl)hex-5-yn-1-yl)oxy]silane

Under argon, intermediate 42 (2.3 g, 10.9 mmol) was dissolved inanhydrous THF (10 mL) at 0° C., and 1.6 M tert-butyllithium in pentane(8.12 mL, 13.0 mmol) was added over a period of 25 min via a syringe,followed by the addition of trimethylsilyl chloride (1.64 mL, 13.0mmol). The resulting suspension was allowed to warm to r.t. and stirredfor 14 h. Et₂O (5 mL) and 0.5 M HCl (2 mL) were added to the reactionmixture, which was then washed with H₂O, 0.5 M HCl, aq. NaHCO₃, andbrine (10 mL). The aqueous layers were extracted with Et₂O (2×30 mL),the combined organic layers were dried over Na₂SO₄ and concentrated invacuo. 43 (2.99 g, 97%) was obtained as a yellow oil, which was usedwithout further purification.

¹H-NMR (500 MHz, CDCl₃): δ 0.04 (s, 6H, TBS-CH₃), 0.13 (s, 9H, TMS-CH₃),0.88 (s, 9H, ^(t)Bu-CH₃), 1.51-1.64 (m, 4H, H-3, H-2), 2.23 (t, J=6.8Hz, 2H, H-4), 3.62 (t, J=6.1 Hz, 2H, H-1); ¹³C-NMR (125 MHz, CDCl₃):δ-5.14 (TBS-CH₃), 0.31 (TMS-CH₃), 18.47 (^(t)Bu-C—CH₃), 19.78 (C-4),25.19 (C-3), 26.10 (^(t)Bu-CH₃), 32.03 (C-2), 62.83 (C-1), 84.59 (C-6),107.52 (C-5).

Synthesis of Intermediate 443-[2-(tert-Butyldimethylsilyloxy)ethyl]-5-nitro-2-(trimethylsilyl)-1H-indole

A mixture of 2-iodo-4-nitroaniline (792 mg, 3.00 mmol), 41 (1.54 g, 6.00mmol), KOAc (1.47 g, 15.0 mmol), LiCl (127 mg, 3.00 mmol) and Pd(OAc)₂(68 mg, 0.3 mmol) in dry DMF (10 mL) was heated at 70-75° C. for 2.5 h.The cooled reaction mixture was diluted with ether (20 mL) and ice-water(20 mL), the aqueous phase was separated and extracted with ethylacetate (3×20 mL). The combined organic phases were washed with waterand brine, dried over Na₂SO₄ and concentrated in vacuo. Purification ofthe residue by chromatography on silica gel (petrol ether/EtOAc 8:1)gave 44 (731 mg, 62%) as a yellow solid.

¹H NMR (500 MHz, CDCl₃): δ −0.01 (s, 6H, 2 CH₃), 0.42 (s, 9H, Si(CH₃)₃),0.85 (s, 9H, C(CH₃)₃), 3.09 (t, J=7.0 Hz, 2H, CH₂), 3.84 (t, J=7.0 Hz,2H, CH₂), 7.36 (d, J=9.0 Hz, 1H, Ar—H), 8.07 (dd, J=2.0, 9.0 Hz, 1H,Ar—H), 8.28 (br s, 1H, Ar—H), 8.62 (d, J=2.5 Hz, 1H, Ar—H); MS: calcdfor C₁₉H₃₂N₂NaO₃Si₂ [M+Na]⁺: 415.18. found 415.10.

Synthesis of Intermediate 453-[3-(tert-Butyldimethylsilyloxy)propyl]-5-nitro-2-(trimethylsilyl)-1H-indole

According to the procedure described for 44, a mixture of2-iodo-4-nitroaniline (1.58 g, 6.00 mmol),tert-butyldimethyl[(5-(trimethylsilyl)pent-4-yn-1-yl)oxy]silane (2.60 g,9.60 mmol), KOAc (2.94 g, 30.0 mmol), LiCl (254 mg, 6.00 mmol) andPd(OAc)₂ (110 mg, 0.6 mmol) in dry DMF (16 mL) was heated at 70-75° C.for 2.5 h. Work-up and purification gave crude 45 (2.40 g) as a yellowsolid.

¹H NMR (500 MHz, CDCl₃): δ 0.1 (s, 6H, 2 CH₃), 0.42 (s, 9H, Si(CH₃)₃),0.94 (s, 9H, C(CH₃)₃), 1.86 (m, 2H, CH₂), 2.93 (t, J=8.0 Hz, 2H, CH₂),3.74 (t, J=6.0 Hz, 2H, CH₂), 7.35 (d, J=9.0 Hz, 1H, Ar—H), 8.07 (dd,J=2.0, 9.0 Hz, 1H, Ar—H), 8.19 (br s, 1H, Ar—H), 8.57 (d, J=1.5 Hz, 1H,Ar—H); ¹³C NMR (125 MHz, CDCl₃): δ −5.3 (2C, 2 CH₃), −0.8 (3C,Si(CH₃)₃), 22.4 (3C, C(CH₃)₃), 26.0 (CH₂), 35.2 (CH₂), 62.7 (CH₂),110.6, 116.4, 117.8, 128.4, 137.1, 141.0, 141.2 (Ar—C).

Synthesis of Intermediate 463-[4-(tert-Butyldimethylsilyloxy)butyl]-5-nitro-2-(trimethylsilyl)-1H-indole

According to the procedure described for 44, a mixture of2-iodo-4-nitroaniline (500 mg, 1.90 mmol), 43 (1.12 g, 3.94 mmol), KOAc(97 mg, 0.988 mmol), LiCl (80 mg, 1.89 mmol) and Pd(OAc)₂ (43 mg, 0.200mmol) in dry DMF (40 mL) was heated at 70-75° C. for 2.5 h. Work-up andpurification gave crude 46 (385 mg, 48%) as a yellow solid.

¹H-NMR (500 MHz, CDCl₃): δ 0.04 (s, 6H, TBS-Me-H), 0.41 (s, 9H,TMS-Me-H), 0.88 (s, 9H, TBS-tBu-H), 1.59-1.68 (m, 2H, H-3′), 1.69-1.75(m, 2H, H-2′), 2.81-2.88 (m, 2H, H-1′), 3.66 (t, J=6.2 Hz, 2H, H-4′),7.34-7.37 (m, 1H, H-7), 8.05-8.08 (m, 1H, H-6), 8.31 (s, 1H, NH),8.55-8.56 (m, 1H, H-4); ¹³C-NMR (125 MHz, CDCl₃): δ −5.16 (TBS-Me-C),−0.60 (TMS-C), 18.43, 26.04, 26.06 (2C, C-1′, TBS-tBu-C), 28.66 (C-2′),33.33 (C-3′), 63.02 (C-4′), 110.79 (C-7), 116.57 (C-4), 117.87 (C-6),128.41, 128.50 (2C, C-3, C-9), 137.27 (C-2), 141.21, 141.36 (2C, C-5,C-8); elemental analysis calcd (%) for C₂₁H₃₆N₂O₃Si₂ (420.69): C, 59.96;H, 8.62; N, 6.66. found: C, 59.85; H, 8.68; N, 6.28.

Synthesis of Intermediate 47 3-(2-Hydroxyethyl)-5-nitro-1H-indole

To a solution of 44 (220 mg, 0.560 mmol) in MeCN (5 mL) was added 48%aq. HF (0.75 mL, 20.7 mmol) in portions. The mixture was stirred at rtfor 48 h, then cautiously basified with saturated aq. Na₂CO₃ andextracted with ethyl acetate. The combined organic phases were washedwith brine, dried over Na₂SO₄ and concentrated. The residue was purifiedby silica gel chromatography (petrol ether/EtOAc 1:1 to 1:3) to afford47 (101 mg, 88%) as yellow solid.

¹H NMR (500 MHz, CD₃OD): δ 3.02 (t, J=7.0 Hz, 2H, CH₂), 3.83 (t, J=7.0Hz, 2H, CH₂), 7.30 (s, 1H, Ar—H), 7.43 (d, J=9.0 Hz, 1H, Ar—H), 8.02(dd, J=2.0, 9.0 Hz, 1H, Ar—H), 8.57 (d, J=2.0 Hz, 1H, Ar—H).

Synthesis of Intermediate 48 3-(3-Hydroxypropyl)-5-nitro-1H-indole

According to the procedure described for 47, to a solution of crude 45(2.40 g) in MeCN (25 mL) was added 48% aq. HF (2.5 mL) and the mixturewas stirred at rt for 48 h. Work-up and purification yielded 48 (820 mg,62% over two steps) as a yellow solid.

¹H NMR (500 MHz, CD₃OD): δ 1.93 (m, 2H, CH₂), 2.86 (t, J=8.0 Hz, 2H,CH₂), 3.64 (t, J=6.0 Hz, 2H, CH₂), 7.24 (s, 1H, Ar—H), 7.41 (d, J=9.0Hz, 1H, Ar—H), 8.00 (dd, J=2.5, 9.0 Hz, 1H, Ar—H), 8.53 (d, J=2.0 Hz,1H, Ar—H); ¹³C NMR (125 MHz, CDCl₃): δ 22.0 (CH₂), 34.3 (CH₂), 62.4(CH₂), 112.2, 116.7, 117.7, 119.1, 126.5, 128.2, 141.2, 142.1 (Ar—C).

Synthesis of Intermediate 49 3-(4-Hydroxybutyl)-5-nitro-1H-indole

To a solution of 46 (371 mg, 0.884 mmol) in anhydrous THF (20 mL) wasadded TBAF (4.4 mL, 1 M in THF) at 0° C. under Argon. The solution wasallowed to warm to r.t., and after 3.5 h, the solvent was removed underreduced pressure. Purification of the residue by silica gelchromatography (CH₂Cl₂, CH₂Cl₂/MeOH 5:1, gradient 0 to 75%) gave 49 (109mg, 53%) as a yellow viscous oil.

¹H-NMR (500 MHz, MeOD): δ 1.59-1.67 (m, 2H, H-3′), 1.74-1.83 (m, 2H,H-2′), 2.78-2.84 (m, 2H, H-1′), 3.60 (t, J=6.5 Hz, 2H, H-4′), 7.23 (s,1H, H-2), 7.38-7.43 (m, 1H, H-7), 7.97-8.02 (m, 1H, H-6), 8.49-8.52 (m,1H, H-4); ¹³C-NMR (125 MHz, MeOD): δ 25.56 (C-1′), 27.78 (C-2′), 33.43(C-3′), 62.80 (C-4′), 112.21 (C-7), 116.70 (C-4), 117.65 (C-6), 119.57(C-3), 126.46 (C-2), 128.18 (C-9), 141.19 (C-8), 142.07 (C-5).

Synthesis of Intermediate 50 3-(2-Bromoethyl)-5-nitro-1H-indole

To a solution of 47 (91.5 mg, 0.443 mmol) in dry MeCN (10 mL) was addedPPh₃ (175 mg, 0.665 mmol) followed by portion-wise addition of CBr₄ (221mg, 0.665 mmol) at 0° C. under argon. The mixture was stirred at rt for2 h, then diluted with ethyl acetate, washed with H₂O and brine anddried over Na₂SO₄. The solvents were evaporated and the residue waspurified by silica gel chromatography (petrol ether/EtOAc 4:1 to 1:1) togive 50 (105 mg, 87%) as a yellow solid.

¹H NMR (500 MHz, DMSO-d6): δ 3.33 (t, J=7.0 Hz, 2H, CH₂), 3.77 (t, J=7.0Hz, 2H, CH₂), 7.52-7.54 (m, 2H, Ar—H), 7.99 (dd, J=2.5, 9.0 Hz, 1H,Ar—H), 8.60 (d, J=2.0 Hz, 1H, Ar—H), 11.71 (s, 1H, Ar—H).

Synthesis of Intermediate 51 3-(3-Bromopropyl)-5-nitro-1H-indole

According to the procedure described for 50, to a solution of 48 (725mg, 3.29 mmol) in MeCN (15 mL) was added PPh₃ (1.30 g, 4.94 mmol) andCBr₄ (1.638 g, 4.94 mmol) at 0° C. under argon. The mixture was stirredat rt for 2 h. Work-up and purification gave 51 (900 mg, 96%) as ayellow solid.

¹H NMR (500 MHz, CD₃OD): δ 2.23 (m, 2H, CH₂), 2.97 (t, J=7.0 Hz, 2H,CH2), 3.48 (t, J=6.5 Hz, 2H, CH₂), 7.29 (s, 1H, Ar—H), 7.44 (d, J=9.0Hz, 1H, Ar—H), 8.03 (dd, J=2.0, 9.0 Hz, 1H, Ar—H), 8.55 (d, J=2.0 Hz,1H, Ar—H); ¹³C NMR (125 MHz, CDCl₃): δ 24.0 (CH₂), 33.9 (CH₂), 34.5(CH₂), 112.4, 116.6, 117.8, 117.8, 127.0, 128.0, 141.2, 142.3 (Ar—C).

Synthesis of Intermediate 52 3-(4-Bromobutyl)-5-nitro-1H-indole

According to the procedure described for 50, to a solution of 49 (100mg, 0.427 mmol) in MeCN (3 mL) was added PPh₃ (135 mg, 0.515 mmol) andCBr₄ (180 mg, 0.543 mmol) at 0° C. under argon. The mixture was stirredat rt for 2 h. Work-up and purification gave 52 (108 mg, 85%) as ayellow solid.

¹H-NMR (500 MHz, CDCl₃): δ 1.85-1.93 (m, 2H, H-2′), 1.93-2.00 (m, 2H,H-3′), 2.83 (t, J=7.4 Hz, 2H, H-1′), 3.46 (t, J=6.6 Hz, 2H, H-4′),7.14-7.18 (m, 1H, H-2), 7.36-7.42 (m, 1H, H-7), 8.08-8.13 (m, 1H, H-6),8.44 (s, 1H, NH), 8.54-8.59 (m, 1H, H-4); ¹³C-NMR (125 MHz, CDCl₃): δ24.17 (C-1′), 28.64 (C-2′), 32.55 (C-3′), 33.68 (C-4′), 111.20 (C-7),116.44 (C-4), 117.89 (C-6), 118.96 (C-3), 124.39 (C-2), 127.08 (C-9),139.45 (C-8), 141.63 (C-5); elemental analysis calcd (%) forC₁₂H₁₃BrN₂O₂ (297.15): C, 48.50; H, 4.41; N, 9.43. found C, 48.55/48.45;H, 4.52/4.54; N, 9.28/9.27.

Synthesis of Product 53 3-(2-Azidoethyl)-5-nitro-1H-indole

To a solution of 50 (240 mg, 0.89 mmol) in dry DMF (7 mL) was added NaN₃(145 mg, 2.23 mmol) and 15-crown-5 (50 μl). The reaction mixture wasstirred at rt for 16 h. The solvent was evaporated and the residue waspurified by silica gel chromatography (petrol ether/EtOAc 3:1) to give53 as yellow solid (202 mg, 98%).

¹H NMR (500 MHz, CDCl₃): δ 3.09 (t, J=7.0 Hz, 2H, CH₂), 3.62 (t, J=7.0Hz, 2H, CH₂), 7.27 (d, J=2.0 Hz, 1H, Ar—H), 7.42 (d, J=9.0 Hz, 1H,Ar—H), 8.13 (dd, J=2.5, 9.0 Hz, 1H, Ar—H), 8.58 (d, J=2.0 Hz, 1H, Ar—H);¹³C NMR (125 MHz, CDCl₃): δ 24.8 (CH₂), 51.4 (CH₂), 111.3, 115.1, 116.0,117.9, 125.4, 126.7, 139.2, 141.7 (Ar—C); IR (KBr): 3409, 2101, 1507,1468, 1326 cm⁻¹; HR-MS Calcd for C₁₀H₉N₅NaO₂ [M+Na]⁺: 254.0654. Found254.0654; elemental analysis calcd (%) for C₁₀H₉N₅O₂: C, 51.95; H, 3.92,N, 30.29. found: C, 52.07; H, 4.04; N, 30.15.

Synthesis of Product 54 3-(3-Azidopropyl)-5-nitro-1H-indole

According to the procedure described for 53, to a solution of 51 (900mg, 3.17 mmol) in DMF (15 mL) was added NaN₃ (309 mg, 4.76 mmol) and15-crown-5 (50 μL). The reaction mixture was stirred at rt for 16 h.Work-up and purification afforded 54 as a yellow solid (725 mg, 93%).

¹H NMR (500 MHz, CDCl₃): δ 2.98 (m, 2H, CH₂), 2.90 (t, J=7.5 Hz, 2H,CH₂), 3.36 (t, J=6.5 Hz, 2H, CH₂), 7.27 (s, 1H, Ar—H), 7.43 (d, J=9.0Hz, 1H, Ar—H), 8.03 (dd, J=2.0, 9.0 Hz, 1H, Ar—H), 8.54 (d, J=2.0 Hz,1H, Ar—H); ¹³C NMR (125 MHz, CDCl₃): δ 22.7 (CH₂), 30.6 (CH₂), 51.9(CH₂), 112.4, 116.6, 117.8, 118.2, 126.8, 128.0, 141.2, 142.2 (Ar—C);HR-MS: calcd for C₁₁H₁₁N₅NaO₂ [M+Na]⁺: 268.0810. found 268.0813.

Synthesis of Product 55 3-(4-Azidobutyl)-5-nitro-1H-indole

According to the procedure described for 53, to a solution of 52 (98 mg,0.330 mmol) in DMF (5 mL) was added NaN₃ (108 mg, 1.66 mmol). Thereaction mixture was stirred at rt for 16 h. Work-up and purificationafforded 55 as a yellow solid (80 mg, 94%).

¹H-NMR (500 MHz, CDCl₃): δ 1.66-1.74 (m, 2H, H-3′), 1.78-1.87 (m, 2H,H-2′), 2.80-2.90 (m, 2H, H-1′), 3.31-3.38 (m, 2H, H-4′), 7.25-7.29 (m,1H, H-2), 7.42-7.47 (m, 1H, H-7), 8.02-8.06 (m, 1H, H-6), 8.53-8.56 (m,1H, H-4); ¹³C-NMR (125 MHz, CDCl₃): δ 25.26 (C-1′), 28.52 (C-2′), 29.66(C-3′), 52.31 (C-4′), 112.28 (C-7), 116.65 (C-4), 117.72 (C-6), 119.15(C-3), 126.56 (C-2), 128.10 (C-9, 141.21 (C-8), 142.14 (C-5); elementalanalysis calcd (%) for C₁₂H₁₃N₅O₂ (259.26): C, 55.59; H, 5.05; N, 27.01.found: C, 55.68/55.75; H, 5.12/5.20; N 26.71/26.73. IR (KBr): ν=3413(s), 2959 (vw), 2920 (w), 2858 (vw), 2090 (vs, N₃), 1619 (w), 1508 (m,NO₂), 1468 (m), 1328 (vs, N₃/NO₂), 1300 (m), 1262 (m), 1212 (w), 1107(w), 1087 (m), 1073 (m), 897 (vw), 814 (w), 733 (w), 584 (vw), 541 (vw)cm⁻¹.

Example 3 Synthesis of the Selectin Antagonists with Triazole Linker(FIG. 6)

General Procedure III for the Synthesis of Antagonists with TriazoleLinker

Alkyne (1.0 equiv.) and azide (1.3 equiv.) were dissolved in^(t)BuOH/H₂O/THF 1:1:1 (approx. 0.015 M) under argon, and the solutionwas degassed in an ultrasound bath for 20 min. Degassed 0.1 M aq.Na-L-ascorbate (0.5 equiv.) and 0.1 M aq. CuSO₄.5H₂O (0.25 equiv.) wereadded, and the mixture was stirred at r.t. for 1-4 h until fullconversion as indicated by MS. The solvent was removed in vacuo, theresidue was redissolved in H₂O/MeCN 1:1+1-2 drops of Et₃N, filtrated(PTFE membrane filter) and purified by preparative HPLC-MS(H₂O/MeCN+0.1% HCOOH). All triazole inhibitors were isolated as yellowsolids.

‡: chemical shift was obtained from HSQC spectrum

§: chemical shift was obtained from HMBC spectrum

Synthesis of Product 56(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-({1-[(6-nitro-1H-indol-3-yl)methyl]-1H-1,2,3-triazol-4-yl}methyl)cyclohexanecarboxamide

Following general procedure III, alkyne 20 (8.4 mg, 10.8 μmol), azide 31(3.6 mg, 16.6 μmol), Na-L-ascorbate (54 μL, 5.40 μmol) and CuSO₄.5H₂O(27 μL, 2.70 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL; noprior degassing). After 3 h, 10.1 mg (94%) of triazole 56 was isolatedby preparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.60 (m, 1H, Cy), 0.61-0.73 (m, 3H, Cy),0.85-0.97 (m, 1H, Cy), 1.06 (d, J=6.4 Hz, 3H, Me), 1.10-1.27 (m, 3H, Cy,H-6_(a), H-2_(a)), 1.26-1.45 (m, 8H, Fuc H-6, 4Cy, Lac H-3_(a)),1.46-1.65 (m, 4H, Lac H-3_(b), H-6_(b), Cy, H-5), 2.07-2.14 (m, 1H,H-2), 2.18-2.26 (m, 1H, H-1), 3.10 (t, J=9.6 Hz, 1H, H-4), 3.54 (t,J=5.8 Hz, 1H, Gal H-5), 3.59-3.68 (m, 2H, Gal H-3, H-3), 3.68-3.80 (m,4H, Fuc H-2, Fuc H-4, 2Gal H-6), 3.85 (dd, J=3.2, 10.3 Hz, 1H, Fuc H-3),3.93-3.97 (m, 1H, Gal H-4), 3.99-4.05 (m, 1H, Lac H-2), 4.30 (t, J=5.3Hz, 2H, H-1′), 4.64 (d, J=8.0 Hz, 1H, Gal H-1), 4.92-5.00 (m, 2H, FucH-1, Fuc H-5), 5.42 (t, J=8.9 Hz, 1H, Gal H-2), 6.77-6.80 (m, 1H, IndH-3), 6.84-6.84 (d, J=14.8 Hz, 1H, H-1″_(a)), 6.88 (d, J=14.8 Hz, 1H,H-1″_(b)), 7.37-7.43 (m, 2H, C₆H₅), 7.52-7.58 (m, 1H, C₆H₅), 7.71-7.74(m, 1H, Ind H-2), 7.86-7.91 (m, 1H, Ind H-7), 8.02 (s, 3H, C₆H₅, H-3′),8.11-8.16 (m, 1H, Ind H-6), 8.19 (t, J=5.4 Hz, 1H, CONH), 8.53-8.56 (m,1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.73 (Fuc C-6), 19.22 (Me),26.55, 26.72, 27.29 (3C, Cy), 33.10, 34.19, 35.00 (3C, Cy), 35.09 (C-2),35.45 (C-1′), 37.48 (C-6), 39.31 (C-5), 42.84 (Lac C-3), 42.95 (C-1),59.70 (C-1″), 62.76 (Gal C-6), 67.71 (2C, Gal C-4, Fuc C-5), 70.28 (FucC-2), 71.39 (Fuc C-3), 72.98 (Gal C-2), 73.95 (Fuc C-4), 75.95 (GalC-5), 78.25 (Lac C-2)‡, 79.70 (C-3), 83.00 (C-4), 83.69 (Gal C-3),100.44 (Fuc C-1), 100.47 (Gal C-1), 106.81 (Ind C-3), 111.48 (Ind C-7),118.74 (Ind C-6), 118.78 (Ind C-4), 123.94 (C-3′), 129.69 (C₆H₅), 130.10(Ind C-9), 130.86 (C₆H₅), 131.52 (C₆H₅), 132.58 (Ind C-2), 134.33(C₆H₅), 140.07 (Ind C-8), 143.86 (Ind C-5), 146.95 (C-2′), 166.85(O(C═O)Ph), 176.93 (CONH), 179.38 (COOH); [a]_(D) ²⁰=−58.0 (c=0.32,MeOH); HPLC (λ=350 nm): purity=95%, t_(R)=14.150 min; IR (KBr): ν=3430(vs, OH), 2927 (s), 2846 (w), 1723 (s, C═O), 1648 (m, C═O), 1616 (w),1584 (vw), 1520 (m), 1476 (vw), 1450 (m), 1403 (w), 1337 (vs, NO₂), 1314(m), 1273 (s), 1226 (w), 1170 (w), 1114 (s), 1092 (s), 1073 (vs), 1040(s), 999 (w), 966 (vw), 766 (w), 746 (w), 713 (m) cm⁻¹. cm⁻¹.

Synthesis of Product 57(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-({1-[2-(5-nitro-1H-indol-1-yl)ethyl]-1H-1,2,3-triazol-4-yl}methyl)cyclohexanecarboxamide

Alkyne 20 (15.0 mg, 19.3 μmol), azide 27 (5.5 mg, 23.8 μmol),Na-L-ascorbate (approx. 1.9 mg, 9.59 μmol), and CuSO₄.5H₂O (1.9 mg, 7.61μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL) under argon (nodegassing). After 2 h, triazole 57 (12.9 mg, 66%) was isolated accordingto general procedure III.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.60 (m, 1H, Cy), 0.61-0.72 (m, 3H, Cy),0.91 (d, J=11.6 Hz, 1H, Me), 1.12-1.25 (m, 3H, Cy, H-6_(a), H-2_(a)),1.25-1.37 (m, 7H, Fuc H-6, 4Cy), 1.37-1.45 (m, 1H, Lac H-3_(a)),1.45-1.61 (m, 3H, Lac H-3_(b), H-6_(b), Cy), 1.61-1.71 (m, 1H, H-5),2.06-2.14 (m, 1H, H-2), 2.20 (tt, J=3.1, 12.8 Hz, 1H, H-1), 3.14 (t,J=9.6 Hz, 1H, H-4), 3.59 (t, J=6.0 Hz, 1H, Gal H-5), 3.64 (dd, J=2.9,9.7 Hz, 1H, Gal H-3), 3.66-3.82 (m, 5H, H-3, Fuc H-2, Fuc H-4, 2GalH-6), 3.87 (dd, J=3.3, 10.3 Hz, 1H, Fuc H-3), 3.94-4.01 (m, 2H, Gal H-4,Lac H-2), 4.15-4.18 (m, 2H), 4.70 (d, J=8.1 Hz, 1H, Gal H-1), 4.75 (t,J=5.3 Hz, 2H, H-2″), 4.78-4.83 (m, 2H, H-1″), 4.97 (t, J=5.1 Hz, 2H, FucH-1, Fuc H-5), 5.43 (dd, J=8.3, 9.5 Hz, 1H, Gal H-2), 6.67-6.69 (m, 1H,Ind H-3), 7.15-7.21 (m, 1H, Ind H-7), 7.21-7.25 (m, 1H, Ind H-2), 7.37(s, 1H, H-3′), 7.38-7.44 (m, 2H, C₆H₅), 7.54-7.60 (m, 1H, C₆H₅),7.96-8.03 (m, 3H, Ind H-6, C₆H₅), 8.06 (t, J=5.3 Hz, 1H, CONH),8.50-8.54 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.74 (Fuc C-6),19.25 (Me), 26.54, 26.69, 27.27 (3C, Cy), 33.11, 34.17 (2C, Cy), 35.05(C-2), 35.09 (C-1′), 35.32 (Cy), 37.51 (C-6), 39.31 (C-5), 42.81 (LacC-3), 42.91 (C-1), 47.53 (C-2″), 51.43 (C-1″), 62.74 (Gal C-6), 67.72(2C, Gal C-4, Fuc C-5), 70.32 (Fuc C-2), 71.41 (Fuc C-3), 72.99 (GalC-2), 73.97 (Fuc C-4), 75.95 (Gal C-5), 78.09 (Lac C-2), 79.78 (C-3),83.01 (C-4), 83.73 (Gal C-3), 100.46 (Fuc C-1), 100.52 (Gal C-1), 105.73(Ind C-3), 110.25 (Ind C-7), 117.99 (Ind C-6), 118.71 (Ind C-4), 124.91(C-3′), 129.18 (Ind C-9), 129.70 (C₆H₅), 130.88 (C₆H₅), 131.54 (C₆H₅),132.55 (Ind C-2), 134.32 (C₆H₅), 140.46 (Ind C-8), 142.89 (Ind C-5),146.55 (C-2′), 166.84 (O(C═O)Ph), 176.74 (CONH), 179.18 (COOH); HR-MS:m/z calcd for C₄₉H₆₄N₆O₁₇ [M+Na]⁺: 1031.4220. found: 1031.4222. HPLC(μ=350 nm): purity=97%, t_(R)=14.483 min; [a]_(D) ²⁰=−53.0 (c=1.09,MeOH); IR (KBr): ν=3435 (vs, OH), 2927 (m), 2852 (w), 1721 (m, C═O),1643 (m, C═O), 1583 (w), 1516 (w), 1449 (w), 1402 (w), 1383 (w), 1335(s, NO₂), 1272 (m). 1215 (vw), 1166 (w), 1111 (m), 1095 (m), 1073 (s),1032 (m), 966 (vw), 897 (vw), 804 (vw), 776 (vw), 765 (vw), 744 (w), 713(w) cm⁻¹.

Synthesis of Product 58(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-({1-[3-(5-nitro-1H-indol-1-yl)propyl]-1H-1,2,3-triazol-4-yl}methyl)cyclohexanecarboxamide

Following general procedure III, alkyne 20 (12.0 mg, 15.4 μmol), azide28 (5.7 mg, 23.2 μmol), Na-L-ascorbate (77 μL, 7.70 μmol), andCuSO₄.5H₂O (39 μL, 3.90 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1(1 mL). After 3 h, triazole 58 (13.5 mg, 85%) was isolated bypreparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.50-0.72 (m, 4H, Cy), 0.85-0.95 (m, 1H, Cy),1.08 (d, J=6.4 Hz, 3H, Me), 1.13-1.25 (m, 3H, H, Cy, H-6_(a), H-2_(a)),1.24-1.37 (m, 7H, 4Cy, Fuc H-6), 1.37-1.44 (m, 1H, Lac H-3_(a)),1.45-1.58 (m, 2H, Lac H-3_(b), Cy), 1.58-1.71 (m, 2H, H-6_(b), H-5),2.13-2.19 (m, 1H, H-2_(b)), 2.25-2.33 (m, 1H, H-1), 2.47 (p, J=6.8 Hz,2H, H-2″), 3.13 (t, J=9.5 Hz, 1H, H-4), 3.56 (t, J=5.8 Hz, 1H, Gal H-5),3.61 (dd, J=2.5, 9.7 Hz, 1H, Gal H-3), 3.73 (qdd, J=5.7, 10.3, 11.7 Hz,5H, H-3, Fuc H-2, Fuc H-4, 2Gal H-6), 3.86 (dd, J=3.3, 10.3 Hz, 1H, FucH-3), 3.95 (s, 1H, Gal H-4), 4.01 (d, J=7.9 Hz, 1H, Lac H-2), 4.31 (t,J=7.0 Hz, 2H, H-3″), 4.33 (s, 2H, H-1′), 4.39 (t, J=6.7 Hz, 2H, H-1″),4.67 (d, J=8.0 Hz, 1H, Gal H-1), 4.93-4.99 (m, 2H, Fuc H-1, Fuc H-5),5.40-5.45 (m, 1H, Gal H-2), 6.72-6.75 (m, 1H, Ind H-3), 7.38-7.45 (m,3H, Ind H-7, C₆H₅), 7.46-7.50 (m, 1H, Ind H-2), 7.53-7.58 (m, 1H, C₆H₅),7.75 (s, 1H, H-3′), 7.99-8.02 (m, 2H, C₆H₅), 8.05-8.09 (m, 1H, Ind H-6),8.20-8.24 (m, 1H, CONH), 8.55-8.57 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz,MeOD): δ 16.73 (Fuc C-6), 19.24 (Me), 26.53, 26.68, 27.26 (3C, Cy),31.48 (C-2″), 33.12, 34.18 (2C, Cy), 35.04, 35.09 (2C, Cy, C-2), 35.64(C-1′), 37.51 (C-6), 39.34 (C-5), 42.77 (Lac C-3), 43.03 (C-1), 44.50(C-3″), 49.85 (C-1″), 62.76 (Gal C-6), 67.72, 67.75 (2C, Gal C-4, FucC-5), 70.31 (Fuc C-2), 71.41 (Fuc C-3), 72.98 (Gal C-2), 73.95 (FucC-4), 75.99 (Gal C-5), 77.98 (Lac C-2)‡, 79.75 (C-3), 83.03 (C-4), 83.71(Gal C-3), 100.43 (Fuc C-1), 100.47 (Gal C-1), 105.24 (Ind C-3), 110.69(Ind C-7), 117.94 (Ind C-6), 118.84 (Ind C-4), 124.44 (C-3′), 129.36(Ind C-9), 129.68 (C₆H₅), 130.87 (C₆H₅), 131.55 (C₆H₅), 132.79 (IndC-2), 134.30 (C₆H₅), 140.24 (Ind C-8), 142.85 (Ind C-5), 166.82(O(C═O)Ph), 176.96 (CONH); HR-MS: m/z calcd for C₅₀H₆₆N₆O₁₇ [M+H]⁺:1023.4557. found: 1023.4561. [a]_(D) ²⁰=−51.9 (c=1.15, MeOH); HPLC(λ=350 nm): purity=96%, t_(R)=14.317 min; IR (KBr): ν=3429 (vs, OH),2928 (m), 2852 (w), 1723 (m, C═O), 1643 (m, CO), 1580 (vw), 1516 (m),1479 (w), 1449 (w), 1402 (w), 1380 (w), 1335 (s, NO₂), 1269 (m), 1210(w), 1163 (w), 1111 (m), 1070 (s), 1032 (m), 963 (w), 894 (vw), 768(vw), 745 (w), 712 (w), 672 (w) cm⁻¹.

Synthesis of Product 59(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-({1-[4-(5-nitro-1H-indol-1-yl)butyl]-1H-1,2,3-triazol-4-yl}methyl)cyclohexanecarboxamide

Following general procedure III, alkyne 20 (9.7 mg, 12.5 μmol), azide 29(4.2 mg, 162 μmol), Na-L-ascorbate (124 μL, 12.4 μmol), and CuSO₄.5H₂O(62 μL, 6.20 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL).After 1 h, triazole 59 (6.4 mg, 49%) was isolated by preparativeHPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.50-0.73 (m, 4H, Cy), 0.86-0.96 (m, 1H, Cy),1.07 (d, J=6.4 Hz, 3H, Me), 1.17 (dt, J=12.5, 35.3 Hz, 3H, Cy, H-6_(a),H-2_(a)), 1.32 (dd, J=8.9, 15.8 Hz, 7H, 4Cy, Fuc H-6), 1.38-1.45 (m, 1H,Lac H-3_(a)), 1.50 (ddd, J=3.9, 10.0, 13.8 Hz, 1H, Lac H-3), 1.54-1.69(m, 3H, Cy, H-6, H-5), 1.77-1.93 (m, 4H, H-3″, H-2″), 2.11-2.18 (m, 1H,H-2), 2.22-2.30 (m, 1H, H-1), 3.12 (t, J=9.5 Hz, 1H, H-4), 3.55 (t,J=5.8 Hz, 1H, Gal H-5), 3.62 (dd, J=2.9, 9.7 Hz, 1H, Gal H-3), 3.64-3.82(m, 5H, H-3, Fuc H-2, Fuc H-4, 2Gal H-6), 3.86 (dd, J=3.3, 10.3 Hz, 1H,Fuc H-3), 3.95-3.97 (m, 1H, Gal H-4), 4.01 (dd, J=2.5, 9.6 Hz, 1H, LacH-2), 4.26 (t, J=6.8 Hz, 2H, H-4″), 4.31 (s, 2H, H-1′), 4.39 (t, J=6.6Hz, 2H, H-1″), 4.67 (d, J=8.1 Hz, 1H, Gal H-1), 4.94-4.99 (m, 2H, FucH-1, Fuc H-5), 5.40-5.45 (m, 1H, Gal H-2), 6.71 (d, J=3.2 Hz, 1H, IndH-3), 7.41-7.47 (m, 3H, Ind H-7, C₆H₅), 7.50-7.54 (m, 1H, Ind H-2),7.56-7.60 (m, 1H, C₆H₅), 7.72 (s, 1H, H-3′), 8.02-8.05 (m, 2H, C₆H₅),8.05-8.09 (m, 1H, Ind H-6), 8.53-8.58 (m, 1H, Ind H-4); ¹³C-NMR (125MHz, MeOD): δ 16.73 (Fuc C-6), 19.23 (Me), 26.54, 26.70, 27.28 (3C, Cy),28.16, 28.48 (2C, C-2″, C-3″), 33.13 (Cy), 34.20 (Cy), 35.06, 35.10 (2C,Cy, C-2), 35.58 (C-1′), 37.44 (C-6), 39.31 (C-5), 42.83 (Lac C-3), 42.97(C-1), 46.81 (C-4″), 50.70 (C-1″), 62.78 (Gal C-6), 67.72, 67.76 (2C,Gal C-4, Fuc C-5), 70.31 (Fuc C-2), 71.41 (Fuc C-3), 73.00 (Gal C-2),73.96 (Fuc C-4), 75.99 (Gal C-5), 78.29 (Lac C-2), 79.76 (C-3), 83.00(C-4), 83.72 (Gal C-3), 100.43 (Fuc C-1), 100.50 (Gal C-1), 104.94 (IndC-3), 110.76 (Ind C-7), 117.80 (Ind C-6), 118.81 (Ind C-4), 124.19(C-3′), 129.27 (Ind C-9), 129.71 (C₆H₅), 130.89 (C₆H₅), 131.59 (C₆H₅),132.79 (Ind C-2), 134.33 (C₆H₅), 140.27 (Ind C-8), 142.72 (Ind C-5),146.16 (C-2′), 166.85 (O(C═O)Ph), 176.90 (CONH), 179.31 (COOH); HR-MS:m/z calcd for C₅₁H₆₈N₆O₁₇ [M−H+2Na]⁺: 1081.4353. found: 1081.4342.[a]_(D) ²⁰=−57.0 (c=0.62, MeOH); HPLC (λ=350 nm): purity=95%,t_(R)=14.633 min; IR (KBr): ν=3431 (vs, OH), 2927 (m), 2852 (m), 1725(m, C═O), 1639 (m, C═O), 1580 (w), 1516 (m), 1479 (w), 1448 (w), 1402(w), 1383 (w), 1334 (s, NO₂), 1271 (m), 1111 (m), 1071 (s), 1032 (m),999 (vw), 966 (vw), 933 (vw), 897 (vw), 837 (vw), 804 (vw), 768 (vw),746 (vw), 713 (w), 667 (vw), 593 (vw) cm⁻¹.

Synthesis of Product 60(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(2-{1-[(6-nitro-1H-indol-3-yl)methyl]-1H-1,2,3-triazol-4-yl}ethyl)cyclohexanecarboxamide

Following general procedure III, alkyne 21 (10.4 mg, 13.1 μmol), azide31 (4.3 mg, 19.8 μmol), Na-L-ascorbate (66 μL, 6.60μmol) and CuSO₄.5H₂O(33 μL, 3.30 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL; noprior degassing). After 4 h, 8.8 mg (66%) of triazole 60 was isolated bypreparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.59 (m, 1H, Cy), 0.59-0.71 (m, 3H, Cy),0.85-0.99 (m, 2H, Cy, H-6_(a)), 1.02 (d, J=6.5 Hz, 3H, Me), 1.04-1.13(m, 1H, H-2_(a)), 1.17-1.37 (m, 8H, Fuc H-6, 5Cy), 1.37-1.45 (m, 2H, LacH-3_(a), H-6), 1.46-1.62 (m, 3H, Lac H-3_(b), H-5, Cy), 2.06-2.12 (m,2H, H-2, H-1), 2.76 (t, J=7.0 Hz, 2H, H-2′), 3.08 (t, J=9.5 Hz, 1H,H-4), 3.55 (t, J=5.8 Hz, 1H, Gal H-5), 3.59-3.66 (m, 2H, Gal H-3, H-3),3.69-3.81 (m, 4H, Fuc H-2, Fuc H-4, 2Gal H-6), 3.86 (dd, J=3.2, 10.3 Hz,1H, Fuc H-3), 3.94-3.97 (m, 1H, Gal H-4), 3.98-4.03 (m, 1H, Lac H-2),4.66 (d, I=8.0 Hz, 1H, Gal H-1), 4.93-5.01 (m, 2H, Fuc H-1, Fuc H-5),5.43 (t, J=8.9 Hz, 1H, Gal H-2), 6.78 (d, J=14.8 Hz, 1H, H-1″_(a)),6.80-6.82 (m, 1H, Ind H-3), 6.85 (d, J=14.7 Hz, 1H, H-1″_(b)), 7.41-7.46(m, 2H, C₆H₅), 7.50-7.55 (m, 1H, C₆H₅), 7.65-7.68 (m, 1H, Ind H-2),7.82-7.85 (m, 1H, H-4′), 7.87-7.90 (m, 1H, Ind H-7), 8.03-8.07 (m, 2H,C₆H₅), 8.14-8.18 (m, 1H, Ind H-6), 8.56-8.59 (m, 1H, Ind H-4); ¹³C-NMR(125 MHz, MeOD): δ 16.74 (Fuc C-6), 19.18 (Me), 26.41, 26.54, 26.72 (3C,2Cy, C-2′), 27.28 (Cy), 33.11, 34.21 35.10 (Cy), 35.35 (C-2′), 37.26(C-6), 39.24 (C-5), 39.66 (C-1′, 42.88, 43.08 (2C, Lac C-3, C-1), 59.63(C-1″), 62.78 (Gal C-6), 67.71 (2C, Gal C-4, Fuc C-5), 70.31 (Fuc C-2),71.42 (Fuc C-3), 73.05 (Gal C-2), 73.98 (Fuc C-4), 76.00 (Gal C-5),78.33 (Lac C-2)‡, 79.87 (C-3), 82.91 (C-4), 83.69 (Gal C-3), 100.44 (FucC-1), 100.62 (Gal C-1), 106.92 (Ind C-3), 111.49 (Ind C-7), 118.78 (IndC-6), 118.84 (Ind C-4), 123.30 (C-4′), 129.73 (C₆H₅), 130.06 (Ind C-9),130.93 (C₆H₅), 131.62 (C₆H₅), 132.52 (Ind C-2), 134.36 (C₆H₅), 140.09(Ind C-8), 143.89 (Ind C-5), 147.06 (C-3′), 166.81 (O(C═O)Ph), 176.99(CONH), 179.82 (COOH)§; [a]_(D) ²⁰=−67.5 (c=0.23, MeOH); HPLC (λ=350nm): purity=95%, t_(R)=14.000 min; IR (KBr): ν=3429 (vs, OH), 2927 (s),2853 (w), 1722 (s, C═O), 1648 (m, C═O), 1616 (w), 1584 (vw), 1520 (m),1476 (vw), 1450 (m), 1403 (w), 1337 (vs, NO₂), 1317 (s), 1273 (s), 1224(w), 1169 (w), 1114 (s), 1095 (s), 1073 (vs), 1034 (s), 1000 (vw), 966(vw), 760 (vw), 746 (w), 713 (m) cm⁻¹.

Synthesis of Product 61(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(2-{1-[2-(5-nitro-1H-indol-1-yl)ethyl]-1H-1,2,3-triazol-4-yl}ethyl)cyclohexanecarboxamide

Alkyne 21 (15.3 mg, 19.3 μmol), azide 27 (5.8 mg, 2.51μmol),Na-L-ascorbate (2.3 mg, 11.6 μmol), and CuSO₄.5H₂O (1.3 mg, 5.21μmol)were dissolved in ^(t)BuOH/H₂O 1:1 (1 mL) under argon (no degassing).After 4 h, triazole 61 (15.7 mg, 79%) was isolated according to generalprocedure III.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.61 (m, 1H, Cy), 0.61-0.73 (m, 3H, Cy),0.83-0.96 (m, 1H, Cy), 1.09 (d, J=6.5 Hz, 3H, Me), 1.11-1.26 (m, 3H,H-2_(a), H-6_(a), Cy), 1.26-1.37 (m, 8H, 5Cy, 3Fuc H-6), 1.37-1.46 (m,1H, Lac H-3_(a)), 1.45-1.61 (m, 3H Lac H-3_(b), H-6, Cy), 1.60-1.70 (m,1H, H-5), 2.06-2.13 (m, 1H, H-2), 2.13-2.22 (m, 1H, H-1), 2.61 (t, J=7.4Hz, 2H, H-2′), 3.09-3.19 (m, 3H, H-1′, H-4), 3.53-3.59 (m, 1H, Gal H-5),3.59-3.83 (m, 6H, Gal H-3, H-3, Fuc H-2, Fuc H-4, 2Gal H-6), 3.87 (dd,J=3.3, 10.3 Hz, 1H, Fuc H-3), 3.91-4.00 (m, 2H, Gal H-4, Lac H-2), 4.68(d, J=8.0 Hz, 1H, Gal H-1), 4.73-4.78 (m, 2H, H-2″), 4.78-4.82 (m, 2H,H-1″), 4.92-5.01 (m, 2H, Fuc H-1, Fuc H-5), 5.39-5.46 (m, 1H, Gal H-2),6.70 (d, J=3.2 Hz, 1H, Ind H-3), 7.24-7.28 (m, 2H, Ind H-2, Ind H-7),7.29 (s, 1H, H-4′), 7.40-7.46 (m, 2H, C₆H₅), 7.51-7.55 (m, 1H, C₆H₅),7.67 (t, J=5.8 Hz, 1H, CONH), 7.97-8.01 (m, 1H, Ind H-6), 8.02-8.06 (m,2H, C₆H₅), 8.49-8.54 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.72(Fuc C-6), 19.25 (Me), 26.35 (C-2′), 26.53, 26.70 (2C, Cy), 27.27 (Cy),33.15 (Cy), 34.22 (Cy), 35.07 (Cy), 35.31 (C-2), 37.38 (C-6), 39.29(C-5), 40.03 (C-1′), 42.82 (Lac C-3), 43.20 (C-1), 47.50 (C-2″), 51.30(C-1″), 62.78 (Gal C-6), 67.71, 67.77 (2C, Gal C-4, Fuc C-5), 70.33 (FucC-2), 71.43 (Fuc C-3), 73.04 (Gal C-2), 73.96 (Fuc C-4), 76.00 (GalC-5), 79.84 (C-3), 82.99 (C-4), 83.66 (Gal C-3), 100.41 (Fuc C-1),100.57 (Gal C-1), 105.73 (Ind C-3), 110.26 (Ind C-7), 117.96 (Ind C-6),118.74 (Ind C-4), 124.34 (C-4′), 129.20 (Ind C-9), 129.69 (C₆H₅), 129.74(C₆H₅), 130.89 (C₆H₅), 132.54 (Ind C-2), 134.32 (C₆H₅), 140.44 (IndC-8), 142.97 (Ind C-5), 146.47 (C-3′), 166.82 (O(C═O)Ph), 176.94 (CONH);HR-MS: m/z calcd for C₅₀H₆₆N₆O₁₇ [M−H+2Na]⁺: 1067.4202. found:1067.4207. HPLC (λ=350 nm): purity=96%, t_(R)=14.500 min; [a]_(D)²⁰=−88.2 (c=0.29, MeOH); IR (KBr): ν=3418 (s, OH), 2926 (s), 2853 (m),1722 (s, C═O), 1648 (m, C═O), 1619 (m), 1583 (w), 1517 (s), 1479 (w),1451 (m), 1404 (w), 1334 (vs, NO₂), 1290 (m), 1271 (w), 1223 (w), 1166(m), 1112 (s), 1095 (s), 1071 (vs), 1032 (s), 967 (w), 999 (m), 935(vw), 897 (vw), 809 (vw), 778 (w), 766 (w), 747 (m), 712 (s), 677 (w),593 (w) cm⁻¹.

Synthesis of Product 62(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(2-{1-[3-(5-nitro-1H-indol-1-yl)propyl]-1H-1,2,3-triazol-4-yl}ethyl)cyclohexanecarboxamide

Alkyne 21 (17.3 mg, 21.8μmol) and azide 28 (7.0 mg, 28.5 μmol) weredissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL). The solution was degassed by3 pump-freeze cycles, and the Na-L-ascorbate (110 μL, 11.0 μmol) andCuSO₄.5H₂O (55 μL, 5.50 μmol) solutions were added under argon. After 3h, triazole 62 (14.4 mg, 63%) was isolated according to generalprocedure III.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.74 (m, 4H, Cy), 0.83-0.97 (m, 1H, Cy),1.05 (d, J=6.5 Hz, 3H, Me), 1.07-1.25 (m, 3H, H-2_(a), H-6_(a, Cy),)1.26-1.37 (m, 7H, Fuc H-6, 4Cy), 1.36-1.45 (m, 1H, Lac H-3_(a)),1.45-1.58 (m, 3H, Lac H-3_(b), H-6_(b, Cy),) 1.58-1.69 (m, 1H, H-5),2.07-2.15 (m, 1H, H-2_(b)), 2.17-2.26 (m, 1H, H-1), 2.46 (p, J=6.6 Hz,2H, H-2″), 2.79 (t, J=7.1 Hz, 2H, H-2′), 3.11 (t, J=9.6 Hz, 1H, H-4),3.32-3.41 (m, 2H, H-1′), 3.54 (t, J=6.1 Hz, 1H, Gal H-5), 3.61 (dd,J=2.8, 9.8 Hz, 1H, Gal H-3), 3.63-3.82 (m, 6H, H-3, Fuc H-1, Fuc H-4,Gal H-6), 3.85 (dd, J=3.3, 10.3 Hz, 1H, Fuc H-3), 3.95 (d, J=2.5 Hz, 1H,Gal H-4), 3.97-4.03 (m, 1H, Lac H-2), 4.30 (t, J=6.9 Hz, 2H, H-3″), 4.37(t, J=6.8 Hz, 2H, H-1″), 4.66 (d, J=8.0 Hz, 1H, Gal H-1), 4.93-5.00 (m,2H, Fuc H-1, Fuc H-5), 5.42 (dd, J=8.2, 9.5 Hz, 1H, Gal H-2), 6.73 (d,J=3.0 Hz, 1H, Ind H-3), 7.41-7.53 (m, 4H, C₆H₆, Ind H-2, Ind H-7), 7.57(m, 1H, C₆H₆), 7.68 (s, 1H, H-4′), 7.81 (t, J=5.3 Hz, 1H, CONH),8.02-8.06 (m, 2H, C₆H₆), 8.06-8.10 (m, 1H, Ind H-6), 8.54-8.58 (m, 1H,Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.72 (Fuc C-6), 19.24 (Me), 26.53,26.62, 26.71 (3C, 2 Cy, C-2′), 27.27 (Cy), 31.48 (C-2″), 33.15 (Cy),34.22 (Cy), 35.07 (Cy), 35.26 (C-2), 37.52 (C-6), 39.31 (C-5), 39.94(C-1′), 42.84 (Lac C-3), 43.23 (C-1), 44.60 (C-3″), [48.16 (C-1″)],62.80 (Gal C-6), 67.72, 67.77 (2C, Gal C-4, Fuc C-5), 70.33 (Fuc C-2),71.42 (Fuc C-3), 73.04 (Gal C-2), 73.96 (Fuc C-4), 76.01 (Gal C-5),[77.78 (Lac C-1)], 79.77 (C-3), 82.98 (C-4), 83.69 (Gal C-3), 100.40(Fuc C-1), 100.52 (Gal C-1), 105.21 (Ind C-3), 110.75 (Ind C-7), 117.92(Ind C-6), 118.83 (Ind C-4), 123.83 (C-4′), 129.39 (Ind C-9), 129.72(C₆H₅), 130.90 (C₆H₅), 131.62 (C₆H₅), 132.84 (Ind C-2), 134.34 (C₆H₅),140.25 (Ind C-8), 142.90 (Ind C-5), [146.71 (C-3′), 166.81 (O(C═O)Ph),177.01 (CONH); HR-MS: m/z calcd for C₅₁H₆₈N₆O₁₇ [—H+2Na]⁺: 1081.4358.found: 1081.4360. HPLC (λ=350 nm): purity=95%, t_(R)=15.033 min; [a]_(D)²⁰=−60.4 (c=0.32, MeOH); IR (KBr): ν=3431 (vs, OH), 2927 (s), 2852 (w),1724 (s, C═O), 1644 (m, C═O), 1578 (w), 1516 (m), 1476 (w), 1451 (m),1404 (w), 1334 (vs, NO₂), 1271 (s), 1220 (w), 1163 (m), 1109 (s), 1071(vs), 1032 (s), 967 (w), 899 (vw), 807 (vw), 765 (w), 746 (m), 713 (m),593 (w) cm⁻¹.

Synthesis of Product 63(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(2-{1-[4-(5-nitro-1H-indol-1-yl)butyl]-1H-1,2,3-triazol-4-yl}ethyl)cyclohexanecarboxamide

Alkyne 21 (14.0 mg, 17.7 μmol), azide 29 (6.1 mg, 23.5 μmol),Na-L-ascorbate (approx. 5 mg, 25.2μmol), and CuSO₄.5H₂O (3.1 mg, 12.5μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL). After 4 h,triazole 63 (9.3 mg, 50%) was isolated according to general procedureIII.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.72 (m, 4H, Cy), 0.84-0.95 (m, 1H, Cy),1.06 (d, J=6.5 Hz, 3H, Me), 1.08-1.26 (m, 3H, Cy, H-6_(a), H-2_(a)),1.26-1.36 (m, 7H, 4 Cy, H-6), 1.36-1.44 (m, 1H, Lac H-3), 1.46-1.59 (m,3H, Lac H-3, H-6_(b), Cy), 1.59-1.66 (m, 1H, H-5), 1.77-1.91 (m, 4H,H-3″, H-2″), 2.09-2.15 (m, 1H, H-2_(b)), 2.19 (tt, J=3.1, 12.6 Hz, 1H,H-1), 2.76 (t, J=7.2 Hz, 2H, H-2′), 3.11 (t, J=9.6 Hz, 1H, H-4), 3.32(m, 1H, H-1′) 3.53-3.58 (m, 1H, Gal H-5), 3.61-3.81 (m, 6H, Gal H-3,H-3, Fuc H-2, Fuc H-4, 2 Gal H-6), 3.86 (dd, J=3.3, 10.3 Hz, 1H, FucH-3), 3.95 (d, J=2.3 Hz, 1H, Gal H-4), 4.03 (dd, J=2.8, 9.8 Hz, 1H, LacH-1), 4.27 (t, J=6.7 Hz, 2H, H-1″), 4.36 (t, J=6.4 Hz, 2H, H-1″), 4.68(d, J=8.1 Hz, 1H, Gal H-1), 4.94 (d, J=4.0 Hz, 1H, Fuc H-1), 4.98 (q,J=6.6 Hz, 1H, Fuc H-5), 5.43 (dd, J=8.3, 9.5 Hz, 1H, Gal H-2), 6.71 (dd,J=0.5, 3.2 Hz, 1H, Ind H—), 7.43-7.47 (m, 3H, C₆H₅, Ind H-2), 7.52-7.58(m, 2H, Ind H-7, C₆H₅), 7.64 (s, 1H, H-4′), 8.03-8.09 (m, 3H, 2 C₆H₅,Ind H-6), 8.56 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.74 (FucC-6), 19.24 (Me), 26.53, 26.59, 26.70 (3C, 2 Cy, C-2′, 27.27 (Cy), 28.11(C-3″), 28.50 (C-2″), 33.12 (Cy), 34.18 (Cy), 35.05 (Cy), 35.27 (C-2),37.46 (C-6), 39.31 (C-5), 39.97 (C-1′), 42.80 (Lac C-3), 43.22 (C-1),46.85 (C-2″), 50.61 (C-1″), 62.75 (Gal C-6), 67.70, 67.74 (2C, Gal C-4,Fuc C-5), 70.31 (Fuc C-2), 71.42 (Fuc C-3), 73.02 (Gal C-2), 73.96 (FucC-4), 75.98 (Gal C-5), 78.07 (Lac C-2), 79.79 (C-3), 83.00 (C-4), 83.68(Gal C-3), 100.43 (Fuc C-1), 100.51 (Gal C-1), 104.93 (Ind C-3), 110.78(Ind C-7), 117.79 (Ind C-6), 118.84 (Ind C-4), 123.58 (C-4′), 129.29(Ind C-9), 129.73 (C₆H₅), 130.91 (C₆H₅), 131.58 (C₆H₅), 132.88 (IndC-2), 134.36 (C₆H₅), 140.24 (Ind C-8), 142.72 (Ind C-5), 146.35 (C-3′),166.81 (O(C═O)Ph), 176.99 (CONH), 179.14 (COOH); HR-MS: m/z calcd forC₅₂H₇₀N₆O₁₇ [M+H]⁺: 1051.4870. found: 1051.4868. HPLC (λ=350 nm):purity=97%, t_(R)=15.283 min; [a]_(D) ²⁰=−60.1 (c=0.86, MeOH); IR (KBr):ν=3414 (vs, OH), 2927 (s), 2852 (m), 1722 (s, C═O), 1648 (m, C═O), 1610(w), 1580 (w), 1515 (m), 1479 (w), 1450 (m), 1404 (w), 1334 (vs), 1272(s), 1215 (w), 1163 (m), 1112 (s), 1072 (vs), 1032 (s, NO₂), 967 (w),897 (vw), 807 (vw), 768 (w), 747 (m), 713 (m), 675 (w), 628 (w), 593 (w)cm⁻¹.

Synthesis of Product 64(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(3-{1-[(5-nitro-1H-indol-1-yl)methyl]-1H-1,2,3-triazol-4-yl}propyl)cyclohexanecarboxamide

Following general procedure III, alkyne 22 (9.7 mg, 12.0 μmol), azide 31(3.9 mg, 18.0 μmol), Na-L-ascorbate (60 μL, 6.00 μmol) and CuSO₄.5H₂O(30 μL, 3.00 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL; noprior degassing). After 3 h, 7.5 mg (61%) of triazole 64 was isolated bypreparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.59 (m, 1H, Cy), 0.59-0.71 (m, 4H, Cy),0.85-0.95 (m, 1H. Cy), 1.09 (d, J=6.5 Hz, 3H, Me), 1.11-1.25 (m, 3H, Cy,H-6_(a), H-2_(a)), 1.25-1.36 (m, 7H, Cy, Fuc H-6), 1.36-1.45 (m, 1H, LacH-3_(a)), 1.45-1.60 (m, 3H, Lac H-3_(b), Cy, H-6_(b)), 1.60-1.67 (m, 1H,H-5), 1.67-1.74 (m, 2H, H-2′), 2.07-2.13 (m, 1H, H-2_(b)), 2.17-2.25 (m,1H, H-1), 2.60 (t, J=7.5 Hz, 2H, H-3′), 3.04-3.10 (m, 2H, H-1′), 3.13(t, J=9.6 Hz, 1H, H-4), 3.56 (t, J=5.9 Hz, 1H, Gal H-5), 3.62-3.81 (m,6H, Gal H-3, H-3, Fuc H-2, Fuc H-4, Gal H-6), 3.86 (dd, J=3.2, 10.3 Hz,1H, Fuc H-3), 3.96 (d, J=2.5 Hz, 1H, Gal H-4), 4.02 (d, J=6.9 Hz, 1H,Lac H-2), 4.66 (d, J=8.0 Hz, 1H, Gal H-1), 4.95-5.00 (m, 2H, Fuc H-1,Fuc H-5), 5.39-5.44 (m, 1H, Gal H-2), 6.79-6.82 (m, 1H, Ind H-3), 6.85(s, 2H, H-1″), 7.24-7.29 (m, 2H, C₆H₅), 7.30-7.35 (m, 1H, C₆H₅),7.69-7.76 (m, 2H, CONH, Ind H-2), 7.88-7.92 (m, 1H, Ind H-7), 7.94 (s,1H, H-5′), 7.96-7.99 (m, 2H, C₆H₅), 8.13-8.17 (m, 1H, Ind H-6),8.55-8.56 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.70 (Fuc C-6),19.24 (Me), 23.46 (C-3′), 26.52 (Cy), 26.70 (Cy), 27.26 (Cy), 30.12(C-2′), 33.12 (Cy), 34.19 (Cy), 35.05 (Cy), 35.52 (C-2), 37.38 (C-6),39.30, 39.42 (2C, C-5, C-1′), 42.78 (Lac C-3), 43.37 (C-1), 59.70(C-1″), 62.77 (Gal C-6), 67.71, 67.76 (2C, Gal C-4, Fuc C-5), 70.33 (FucC-2), 71.42 (Fuc C-3), 73.04 (Gal C-2), 73.97 (Fuc C-4), 75.99 (GalC-5), 78.28 (Lac C-2)‡, 79.91 (C-3), 82.98 (C-4), 83.61 (Gal C-3),100.45 (Fuc C-1), 100.63 (Gal C-1), 106.81 (Ind C-3), 111.47 (Ind C-7),118.80, 118.81 (2C, Ind C-4, Ind C-6), 123.10 (C-5′), 129.58 (Ind C-9),130.11 (C₆H₅), 130.76 (C₆H₅), 131.48 (C₆H₅), 132.58 (Ind C-2), 134.21(C₆H₅), 140.11 (Ind C-8), 143.89 (Ind C-5), 149.16 (C-4′), 166.76(O(C═O)Ph), 177.05 (CONH); HR-MS: m/z calcd for C₅₀H₆₆N₆O₁₇ [M+Na]⁺:1045.4377. found: 1045.4376; [a]_(D) ²⁰=−42.3 (c=0.72, MeOH).

Synthesis of Product 65(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(3-{1-[2-(5-nitro-1H-indol-1-yl)ethyl]-1H-1,2,3-triazol-4-yl}propyl)cyclohexanecarboxamide

Following general procedure III, alkyne 22 (14.6 mg, 18.1 μmol), azide27 (5.5 mg, 23.8 μmol), Na-L-ascorbate (91 μL, 9.10 μmol) and CuSO₄.5H₂O(45 μL, 4.50 μmol) were dissolved in ^(t)BuOH/H₂O/MeCN 1:1 (0.8 mL) andMeCN (0.1 mL). As no product could be detected after 1 h, further 91 μLand 45 μL of the Na-L-ascorbate and CuSO₄.5H₂O solution were added.After another 18 h, triazole 65 (12.8 mg, 68%) was isolated bypreparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.48-0.59 (m, 1H, Cy), 0.59-0.70 (m, 3H, Cy),0.84-0.93 (m, 1H, Cy), 1.11 (d, J=6.5 Hz, 3H, Me), 1.14-1.36 (m, 10H,Cy, H-6_(a), H-2_(a), 4Cy, Fuc H-6), 1.36-1.44 (m, 1H, Lac H-3_(a)),1.44-1.54 (m, 3H, Lac H-3_(b), H-2′), 1.54-1.62 (m, 2H, Cy, H-6_(b)),1.62-1.72 (m, 1H, H-5), 2.12-2.18 (m, 1H, H-2_(b)), 2.19-2.27 (m, 1H,H-1), 2.47 (t, J=7.4 Hz, 2H, H-3′), 2.86-2.93 (m, 2H, H-1′), 3.15 (t,J=9.6 Hz, 1H, H-4), 3.59 (t, J=5.8 Hz, 1H, Gal H-5), 3.65 (dd, J=2.7,9.7 Hz, 1H, Gal H-3), 3.67-3.82 (m, 5H, H-3, Fuc H-2, Fuc H-4, 2GalH-6), 3.88 (dd, J=3.3, 10.3 Hz, 1H, Fuc H-3), 3.94-3.99 (m, 2H, Gal H-4,Lac H-2), 4.72-4.78 (m, 3H, Gal H-1, H-2″), 4.79-4.83 (m, 2H, H-1″),4.95-5.02 (m, 2H, Fuc H-1, Fuc H-5), 5.43 (dd, J=8.4, 9.3 Hz, 1H, GalH-2), 6.69-6.71 (m, 1H, Ind H-3), 7.15-7.19 (m, 1H, Ind H-7), 7.21 (s,1H, H-5′), 7.31-7.34 (m, 1H, Ind H-2), 7.35-7.39 (m, 2H, C₆H₅),7.49-7.54 (m, 1H, C₆H₅), 7.94-7.97 (m, 1H, Ind H-6), 8.01-8.04 (m, 2H,C₆H₅), 8.48-8.52 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.73 (FucC-6), 19.27 (Me), 23.28 (C-3′), 26.54, 26.71, 27.30 (3C, Cy), 30.26(C-2′), 33.11, 34.24, 35.13 (3C, Cy), 35.38 (C-2), 37.49 (C-6), 39.31,39.36 (2C, C-5, C-1′), 42.98 (Lac C-3), 43.32 (C-1), 47.51 (C-2″), 51.46(C-1″), 62.87 (Gal C-6), 67.73, 67.76 (2C, Gal C-4, Fuc C-5), 70.34 (FucC-2), 71.42 (Fuc C-3), 73.02 (Gal C-2), 73.97 (Fuc C-4), 76.03 (GalC-5), 78.78 (Lac C-2)¹, 79.85 (C-3), 83.05 (C-4), 83.72 (Gal C-3),100.44 (Fuc C-1), 100.56 (Gal C-1), 105.87 (Ind C-3), 110.18 (Ind C-7),117.98 (Ind C-6), 118.73 (Ind C-4), 124.12 (C-5′), 129.11 (Ind C-9),129.65 (C₆H₅), 130.85 (C₆H₅), 131.62 (C₆H₅), 132.46 (Ind C-2), 134.25(C₆H₅), 140.57 (Ind C-8), 142.88 (Ind C-5), 148.44 (C-4′), 166.82(O(C═O)Ph), 176.99 (CONH), 180.71 (COOH); HR-MS: m/z calcd forC₅₁H₆₈N₆O₁₇ [M+H]⁺: 1037.4714. found: 1037.4711. HPLC (λ=350 nm):purity=96%, t_(R)=14.583 min; [α]_(D)=−51.0 (c=0.89, MeOH); IR (KBr):ν=3421 (s, OH), 2927 (m), 2852 (w), 1718 (m, C═O), 1646 (m, C═O), 1638(m), 1620 (m), 1580 (w), 1517 (m), 1476 (w), 1429 (w), 1402 (w), 1383(w), 1336 (vs, NO₂), 1270 (m), 1218 (w), 1166 (w), 1095 (s), 1070 (s),1029 (s), 996 (m), 966 (w), 933 (vw), 894 (vw), 826 (vw), 809 (vw), 779(vw), 771 (vw), 746 (w), 712 (w), 675 (vw), 589 (vw) cm⁻¹.

Synthesis of Product 66(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(3-{1-[3-(5-nitro-1H-indol-1-yl)propyl]-1H-1,2,3-triazol-4-yl}propyl)cyclohexanecarboxamide

Alkyne 22 (106 mg, 132 μmol) and azide 28 (48.0 mg, 196 μmol) weredissolved in ^(t)BuOH/H₂O/THF 1:1:1 (8 mL), and the 0.1 M Na-L-ascorbate(658 μL, 65.8 μmol) and CuSO₄.5H₂O (329 μL, 32.9μmol) solutions wereadded under argon. After 40 min, the THF was removed under reducedpressure, and the residue was subjected to RP-18 silica gelchromatography (MeOH in H₂O, linear gradient, 0 to 95%). The productfractions were concentrated and further purified by preparative HPLC-MS(H₂O/MeCN+0.1% HCOOH) to afford triazole 66 (125 mg, 91%).

¹H-NMR (500 MHz, MeOD): δ 0.46-0.70 (m, 4H, Cy), 0.81-0.93 (m, 1H, Cy),1.11 (d, J=6.5 Hz, 3H, Me), 1.14-1.36 (m, 10H, H-2_(a), H-6_(a), 5Cy,Fuc H-6), 1.36-1.44 (m, 1H, Lac H-3_(a)), 1.44-1.50 (m, 1H, LacH-3_(b)), 1.50-1.57 (m, 1H, Cy), 1.60 (dd, J=2.5, 13.2 Hz, 1H, H-6_(b)),1.63-1.71 (m, 1H H-5), 1.71-1.81 (m, 2H, H-2′), 2.10-2.19 (m, 1H,H-2_(b)), 2.25 (ddd, J=3.2, 8.0, 12.7 Hz, 1H, H-1), 2.49 (p, J=6.8 Hz,2H, H-2″), 2.65 (t, J=7.4 Hz, 2H, H-3′), 3.05-3.19 (m, 3H, H-1′, H-4),3.57 (t, J=6.1 Hz, 1H, Gal H-5), 3.64 (dd, J=3.0, 9.7 Hz, 1H, Gal H-3),3.66-3.83 (m, 5H, H-3, Fuc H-2, Fuc H-4, Gal H-6), 3.87 (dd, J=3.3, 10.3Hz, 1H, Fuc H-3), 3.96 (d, J=2.4 Hz, 1H, Gal H-4), 4.00 (dd, J=2.9, 9.8Hz, 1H, Lac H-2), 4.32 (t, J=6.9 Hz, 2H, H-3″), 4.40 (t, J=6.7 Hz, 2H,H-1″), 4.70 (d, J=8.1 Hz, 1H, Gal H-1), 4.95-5.01 (m, 2H, Fuc H-1, FucH-5), 5.43 (dd, J=8.2, 9.6 Hz, 1H, Gal H-2), 6.71-6.73 (m, 1H, Ind H-3),7.34-7.38 (m, 2H, C₆H₅), 7.44-7.52 (m, 3H, Ind H-7, Ind H-2, C₆H₅), 7.65(s, 1H, H-5′), 7.98-8.02 (m, 2H, C₆H₅), 8.04-8.08 (m, 1H, Ind H-6),8.55-8.56 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.73 (Fuc C-6),19.26 (Me), 23.54 (C-3′), 26.50, 26.67, 27.25 (3C, Cy), 30.22 (C-2′),31.41 (C-2″), 33.11, 34.18, 35.04 (3C, Cy), 35.42 (C-2), 37.50 (C-6),39.35, 39.48 (2C, C-5, C-1′), 42.82 (Lac C-3), 43.35 (C-1), 44.64(C-3″), 48.56 (C-1″), 62.79 (Gal C-6), 67.71, 67.77 (2C, Gal C-4, FucC-5), 70.33 (Fuc C-2), 71.42 (Fuc C-3), 73.00 (Gal C-2), 73.96 (FucC-4), 75.99 (Gal C-5), 78.32 (Lac C-2), 79.85 (C-3), 83.03 (C-4), 83.67(Gal C-3), 100.45 (Fuc C-1), 100.54 (Gal C-1), 105.21 (Ind C-3), 110.70(Ind C-7), 117.90 (Ind C-6), 118.83 (Ind C-4), 123.54 (C-5′), 129.36(Ind C-9), 129.65 (C₆H₅), 130.83 (C₆H₅), 131.56 (C₆H₅), 132.81 (IndC-2), 134.26 (C₆H₅), 140.24 (Ind C-8), 142.83 (Ind C-5), 148.51 (C-4′),166.78 (O(C═O)Ph), 177.04 (CONH), 179.26 (COOH); HR-MS: m/z calcd forC₅₂H₇₀N₆O₁₇ [M+Na]⁺: 1073.4690. found: 1073.4684. [a]_(D) ²⁰=−53.1(c=0.99, MeOH); HPLC (λ=350 nm): purity=97%, t_(R)=14.267 min; IR (KBr):ν=3433 (vs, OH), 2926 (m), 2852 (w), 1725 (m, C═O), 1638 (m, C═O), 1514(w), 1449 (w), 1402 (w), 1383 (w), 1334 (m, NO₂), 1268 (m), 1213 (w),1163 (w), 1111 (m), 1070 (s), 1032 (m), 996 (w), 963 (vw), 933 (vw), 897(vw), 807 (vw), 777 (vw), 744 (w), 712 (w) cm⁻¹.

Synthesis of Product 67(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(3-{1-[4-(5-nitro-1H-indol-1-yl)butyl]-1H-1,2,3-triazol-4-yl}propyl)cyclohexanecarboxamide

Following general procedure III, alkyne 21 (12.0 mg, 14.9 μmol) andazide 29 (5.8 mg, 22.4 μmol), Na-L-ascorbate (74 μL, 7.40 μmol), andCuSO₄.5H₂O (36 μL, 3.70 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1(1 mL). After 2 h, triazole 67 (9.4 mg, 59%) was isolated by preparativeHPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.70 (m, 4H, Cy), 0.83-0.94 (m, 1H, Cy),1.10 (d, J=6.4 Hz, 3H, Me), 1.14-1.37 (m, 10H, H-2_(a), H-6_(a, Cy,)4Cy, Fuc H-6), 1.40 (dd, J=9.1, 11.8 Hz, 1H, Lac H-3_(a)), 1.45-1.62 (m,3H, Lac H-3_(b), Cy, H-6_(b)), 1.62-1.70 (m, 1H, H-5), 1.70-1.78 (m, 2H,H-2′), 1.78-1.93 (m, 4H, H-3″, H-2″), 2.11-2.18 (m, 1H, H-2_(b)),2.21-2.29 (m, 1H, H-1), 2.63 (t, J=7.4 Hz, 2H, H-3′), 3.03-3.19 (m, 3H,H-1′, H-4), 3.57 (t, J=5.9 Hz, 1H, Gal H-5), 3.64 (dd, J=2.8, 9.7 Hz,1H, Gal H-3), 3.66-3.83 (m, 5H, H-3, Fuc H-2, Fuc H-4, Gal H-6), 3.87(dd, J=3.2, 10.3 Hz, 1H, Fuc H-3), 3.95-4.03 (m, 2H, Gal H-4, Lac H-2),4.28 (t, J=6.8 Hz, 2H, H-4″), 4.38 (t, J=6.6 Hz, 2H, H-1″), 4.70 (d,J=8.0 Hz, 1H, Gal H-1), 4.95-5.02 (m, 2H, Fuc H-1, Fuc H-5), 5.40-5.45(m, 1H, Gal H-2), 6.68-6.72 (m, J=2.8 Hz, 1H, Ind H-3), 7.33-7.40 (m,2H, C₆H₅), 7.42-7.46 (m, 1H, Ind H-2), 7.46-7.54 (m, 2H, C₆H₅, Ind H-7),7.63 (s, 1H, H-5′), 7.98-8.04 (m, 2H, C₆H₅), 8.04-8.08 (m, 1H, Ind H-6),8.53-8.57 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.73 (Fuc C-6),19.26 (Me), 23.53 (C-3′), 26.52, 26.69, 27.27 (3C, Cy), 28.11, 28.47(2C, C-2″, C-3″), 30.23 (C-2′), 33.12, 34.20, 35.07 (3C, Cy), 35.44(C-2), 37.48 (C-6), 39.34, 39.45 (2C, C-5, C-1′), 42.86 (2C, C-5, C-1′),43.34 (C-1), 46.87 (C-2″), 50.61 (C-1″), 62.81 (Gal C-6), 67.71, 67.76(2C, Gal C-4, Fuc C-5), 70.33 (Fuc C-2), 71.42 (Fuc C-3), 73.02 (GalC-2), 73.97 (Fuc C-4), 76.00 (Gal C-5), 78.36 (Lac C-2), 79.87 (C-3),83.01 (C-4), 83.67 (Gal C-3), 100.44 (Fuc C-1), 100.58 (Gal C-1), 104.90(Ind C-3), 110.78 (Ind C-7), 117.78 (Ind C-6), 118.82 (Ind C-4), 123.35(C-5′), 129.31 (Ind C-9), 129.65 (C₆H₅), 130.83 (C₆H₅), 131.57 (C₆H₅),132.89 (Ind C-2), 134.26 (C₆H₅), 140.26 (Ind C-8), 142.71 (Ind C-5),148.45 (C-4′), 166.78 (O(C═O)Ph), 177.03 (CONH), 179.61 (COOH); HR-MS:m/z calcd for C₅₃H₇₂N₆O₁₇ [M+Na]⁺: 1087.4846. found: 1087.4848. [a]_(D)²⁰=−53.1° (c=0.82, MeOH); HPLC (λ=350 nm): purity=99%, t_(R)=14.517 min;IR (KBr): ν=3436 (vs, OH), 2926 (w), 2852 (vw), 1723 (w, C═O), 1632 (m,C═O), 1512 (w), 1451 (w), 1383 (w), 1334 (m, NO₂), 1270 (m), 1210 (vw),1166 (vw), 1111 (m), 1070 (m), 1032 (m), 996 (vw), 966 (vw), 744 (w),711 (w) cm⁻¹.

Synthesis of Product 68(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(4-{1-[(6-nitro-1H-indol-3-yl)methyl]-1H-1,2,3-triazol-4-yl}butyl)cyclohexanecarboxamide

Following general procedure III, alkyne 23 (10.6 mg, 13.4μmol), azide 31(4.4 mg, 20.3 μmol), Na-L-ascorbate (67 μL, 6.70 μmol) and CuSO₄.5H₂O(34 μL, 3.40 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL; noprior degassing). After 2 h, 11.3 mg (81%) of triazole 68 was isolatedby preparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.47-0.70 (m, 4H, Cy), 0.83-0.94 (m, 1H, Cy),1.08 (d, J=6.4 Hz, 3H, Me), 1.11-1.24 (m, 3H, Cy, H-6_(a), H-2_(a)),1.25-1.32 (m, 4H, Cy), 1.34 (d, J=6.5 Hz, 3H, Fuc H-6), 1.37-1.68 (m,9H, Lac H-3_(a), Lac H-3_(b), H-2′, H-6, H-3′, H-5, Cy), 2.06-2.13 (m,1H, H-2_(b)), 2.16-2.24 (m, 1H, H-1), 2.66 (t, J=7.5 Hz, 2H, H-4′), 3.06(t, J=7.0 Hz, 2H, H-1′), 3.13 (t, J=9.6 Hz, 1H, H-4), 3.58 (t, J=5.9 Hz,1H, Gal H-5), 3.63-3.70 (m, 2H, Gal H-3, H-3), 3.75 (s, 4H, Fuc H-2, FucH-4, 2Gal H-6), 3.86 (dd, J=3.2, 10.3 Hz, 1H, Fuc H-3), 3.96 (d, J=2.0Hz, 1H, Gal H-4), 4.02 (dd, J=2.5, 9.7 Hz, 1H, Lac H-2), 4.69 (d, J=8.0Hz, 1H, Gal H-1), 4.95-5.02 (m, 3H, Fuc H-1, Fuc H-5), 5.43 (t, J=8.9Hz, 1H, Gal H-2), 6.78-6.81 (m, 1H, Ind H-3), 6.83 (s, 2H, H-1″),7.36-7.41 (m, 2H, C₆H₅), 7.51-7.56 (m, 1H, C₆H₅), 7.70-7.74 (m, 1H, IndH-2), 7.86-7.90 (m, 1H, Ind H-7), 7.95 (s, 1H, H-6′), 7.99-8.03 (m, 2H,C₆H₅), 8.12-8.17 (m, 1H, Ind H-6), 8.54-8.57 (m, 1H, Ind H-4); ¹³C-NMR(125 MHz, MeOD): δ 16.74 (Fuc C-6), 19.24 (Me), 25.67 (C-4′), 26.51,26.67, 27.24 (3C, Cy), 27.43 (C-3′), 29.67 (C-2′), 33.08 (Cy), 34.13(Cy), 35.03 (Cy), 35.38 (C-2), 37.46 (C-6), 39.32 (C-5), 39.75 (C-1′),42.75 (Lac C-3), 43.31 (C-1), 59.65 (C-1″), 62.73 (Gal C-6), 67.70,67.75 (2C, Gal C-4, Fuc C-5), 70.30 (Fuc C-2), 71.41 (Fuc C-3), 73.00(Gal C-2), 73.96 (Fuc C-4), 75.95 (Gal C-5), 77.96 (Lac C-2), 79.82(C-3), 83.02 (C-4), 83.63 (Gal C-3), 100.45 (Fuc C-1), 100.50 (Gal C-1),106.78 (Ind C-3), 111.43 (Ind C-7), 118.77 (Ind C-6), 118.82 (Ind C-4),122.98 (C-6′), 129.65 (C₆H₅), 130.07 (Ind C-9), 130.84 (C₆H₅), 131.52(C₆H₅), 132.59 (Ind C-2), 134.34 (C₆H₅), 140.07 (Ind C-8), 143.83 (IndC-5), 149.56 (C-5′), 166.79 (O(C═O)Ph), 176.95 (CONH), 178.88 (COOH);[a]_(D) ²⁰=−53.2 (c=1.13, MeOH); HPLC (λ=350 nm): purity=96%,t_(R)=14.367 min; IR (KBr): ν=3436 (vs, OH), 2927 (s), 2852 (w), 1725(m, C═O), 1647 (m, C═O), 1616 (w), 1583 (vw), 1520 (m), 1476 (vw), 1450(m), 1402 (w), 1384 (w), 1336 (s, NO₂), 1315 (m), 1271 (m), 1223 (w),1167 (w), 1114 (m), 1072 (s), 1040 (s), 999 (w), 966 (vw), 763 (vw), 746(w), 712 (w) cm⁻¹.

Synthesis of Product 69(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(4-{1-[2-(6-nitro-1H-indol-3-yl)ethyl]-1H-1,2,3-triazol-4-yl}butyl)cyclohexanecarboxamide

Following general procedure III, alkyne 23 (10.2 mg, 12.9 μmol), azide27 (4.5 mg, 19.5 μmol), Na-L-ascorbate (64 μL, 6.40 μmol) and CuSO₄.5H₂O(32 μL, 3.20 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL; noprior degassing). After 2.5 h, 11.5 mg (85%) of triazole 69 was isolatedby preparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.49-0.59 (m, 1H, Cy), 0.59-0.71 (m, 3H, Cy),0.85-0.95 (m, 1H, Cy), 1.10 (d, J=6.5 Hz, 3H, Me), 1.14-1.24 (m, 3H, Cy,H-6_(a), H-2_(a)), 1.24-1.36 (m, 9H, Fuc H-6, 4Cy, H-2′), 1.36-1.61 (m,6H, H-3′, Lac H-3_(b), H-6_(b), Cy), 1.61-1.71 (m, 1H, H-5), 2.10-2.16(m, 1H, H-2), 2.20-2.28 (m, 1H, H-1), 2.50 (t, J=7.3 Hz, 2H, H-4′),2.96-3.07 (m, 2H, H-1′), 3.14 (t, J=9.6 Hz, 1H, H-4), 3.57 (t, J=5.7 Hz,1H, Gal H-5), 3.64 (dd, J=2.5, 9.7 Hz, 1H, Gal H-3), 3.66-3.82 (m, 5H,H-3, Fuc H-2, Fuc H-4, Gal H-6), 3.87 (dd, J=3.2, 10.3 Hz, 1H, Fuc H-3),3.94-3.97 (m, 1H, Gal H-4), 4.00-4.05 (m, 1H, Lac H-2), 4.71 (d, J=8.0Hz, 1H, Gal H-1), 4.74-4.78 (m, 2H, H-2″), 4.79-4.83 (m, 2H, H-1″),4.95-5.01 (m, 2H, Fuc H-1, Fuc H-5), 5.40-5.46 (m, 1H, Gal H-2),6.69-6.72 (m, 1H, Ind H-3), 7.19-7.22 (m, 1H, Ind H-7), 7.22 (s, 1H,H-6′), 7.27-7.30 (m, 1H, Ind H-2), 7.41-7.46 (m, 2H, C₆H₅), 7.55-7.60(m, 1H, C₆H₅), 7.95-7.99 (m, 1H, Ind H-6), 8.01-8.04 (m, 2H, C₆H₅), 8.52(s, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.72 (Fuc C-6), 19.24(Me), 25.46 (C-4′), 26.49 (Cy), 26.66 (Cy), 27.24 (Cy), 27.59 (C-3′),29.51 (C-2′), 33.08 (Cy), 34.13 (Cy), 35.02 (Cy), 35.32 (C-2), 37.52(C-6), 39.34 (C-5), 39.79 (C-1′), 42.76 (Lac C-3), 43.27 (C-1), 47.49(C-2″), 51.34 (C-1″), 62.72 (Gal C-6), 67.68 (Gal C-4), 67.73 (Fuc C-5),70.29 (Fuc C-2), 71.39 (Fuc C-3), 72.95 (Gal C-2), 73.94 (Fuc C-4),75.94 (Gal C-5), 77.98 (Lac C-2), 79.81 (C-3), 83.03 (C-4), 83.65 (GalC-3), 100.44 (Fuc C-1), 100.47 (Gal C-1), 105.76 (Ind C-3), 110.21 (IndC-7), 117.97 (Ind C-6), 118.69 82 (Ind C-4), 123.96 (C-6′), 129.12 (IndC-9), 129.68 (C₆H₅), 130.84 (C₆H₅), 131.51 (C₆H₅), 132.46 (Ind C-2),134.34 (C₆H₅), 140.49 (Ind C-8), 142.87 (Ind C-5), 148.89 (C-5′), 166.79(O(C═O)Ph), 176.92 (CONH), 178.93 (COOH); [a]_(D) ²⁰=−47.8 (c=1.10,MeOH); HPLC (λ=350 nm): purity=97%, t_(R)=14.000 min; IR (KBr): ν=3436(vs, OH), 2927 (m), 2852 (w), 1725 (m, C═O), 1638 (m, C═O), 1584 (vw),1516 (w), 1479 (vw), 1450 (w), 1383 (vw), 1335 (s, NO₂), 1269 (s), 1218(w), 1166 (w), 1114 (m), 1071 (s), 1032 (m), 998 (w), 966 (vw), 746(vw), 712 (w) cm⁻¹.

Synthesis of Product 70(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-({1-[2-(6-nitro-1H-indol-3-yl)ethyl]-1H-1,2,3-triazol-4-yl}methyl)cyclohexanecarboxamide

Following general procedure III, alkyne 20 (9.2 mg, 11.8 μmol), azide 53(3.6 mg, 15.6 μmol), Na-L-ascorbate (118 μL, 11.8 μmol), and CuSO₄.5H₂O(59 μL, 5.90 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL).After 1 h, the solvent was removed and the residue was purified by threepreparative HPLC-MS runs (H₂O/MeCN+0.1% HCOOH). After the firstpurification, 5.7 mg (48%; purity<90%) of triazole 70 were obtained,after the third purification, 2.5 mg (21%; purity=99%) were isolated.

¹H-NMR (500 MHz, MeOD): δ 0.48-0.71 (m, 4H, 4 Cy), 0.84-0.95 (m, 1H,Cy), 1.10 (t, J=7.0 Hz, 3H, Me), 1.13-1.24 (m, 3H, H-2_(a), H-6_(a),Cy), 1.24-1.36 (m, 7H, 4Cy, Fuc H-6), 1.36-1.45 (m, 1H, Lac H-3_(a)),1.45-1.53 (m, 1H, Lac H-3_(b)), 1.53-1.61 (m, 2H, Cy, H-6_(b)),1.60-1.70 (m, 1H, H-5), 2.11-2.18 (m, 1H, H-2_(b)), 2.20-2.29 (m, 1H,H-1), 3.14 (t, J=9.6 Hz, 1H, H-4), 3.41 (t, J=6.7 Hz, 2H, H-2″), 3.57(t, J=5.9 Hz, 1H, Gal H-5), 3.61 (dd, J=2.9, 9.7 Hz, 1H, Gal H-3),3.65-3.82 (m, 5H, H-3, Fuc H-2, Fuc H-4, Gal H-6), 3.87 (dd, J=3.3, 10.3Hz, 1H, Fuc H-3), 3.93-3.98 (m, 2H, Gal H-4, Lac H-2), 4.27 (s, 2H,H-1′), 4.66-4.71 (m, 3H, H-1″, Gal H-1), 4.94-5.00 (m, 2H, Fuc H-1, FucH-5), 5.42 (dd, J=8.2, 9.6 Hz, 1H, Gal H-2), 7.21 (s, 1H, Ind H-2),7.41-7.46 (m, 3H, C₆H₅, Ind H-7), 7.55-7.59 (m, 1H, C₆H₅), 7.66 (s, 1H,H-3′), 8.01-8.05 (m, 3H, Ind H-6, C₆H₅), 8.29-8.31 (m, 1H, Ind H-4);¹³C-NMR (125 MHz, MeOD): δ 16.72 (Fuc C-6), 19.26 (Me), 26.54, 26.72,27.09 (3C, Cy), 27.30 (C-2″), 33.12 (Cy), 34.24 (Cy), 35.12, 35.16 (2C,Cy, C-2), 35.54 (C-1′), 37.39 (C-6), 39.32 (C-5), 42.95, 42.98 (2C, C-1,Lac C-3), 52.36 (C-1″), 62.85 (Gal C-6), 67.74, 67.75 (2C, Gal C-4, FucC-5), 70.33 (Fuc C-2), 71.41 (Fuc C-3), 73.02 (Gal C-2), 73.96 (FucC-4), 76.03 (Gal C-5), 79.11 (Lac C-2)‡, 79.79 (C-3), 83.03 (C-4), 83.75(Gal C-3), 100.44 (Fuc C-1), 100.54 (Gal C-1), 112.57 (Ind C-7), 114.78(Ind C-3), 116.40 (Ind C-4), 117.99 (Ind C-6), 124.51 (C-3′), 127.90,127.98 (2C, Ind C-2, Ind C-9), 129.69 (C₆H₅), 130.89 (C₆H₅), 131.60(C₆H₅), 134.30 (C₆H₅), 141.03 (Ind C-8), 142.51 (Ind C-5), 146.17(C-2′)‡, 166.86 (O(C═O)Ph), 176.85 (CONH); HR-MS: m/z calcd forC₄₉H₆₄N₆O₁₇ [M+Na]⁺: 1031.4220. found: 1031.4223. [a]_(D) ²⁰=−62.7(c=0.26, MeOH); HPLC (λ=350 nm): purity=99%, t_(R)=13.517 min; IR (KBr):ν=3435 (vs, OH), 2925 (w), 2852 (vw), 1720 (w, C═O), 1632 (m, C═O), 1517(vw), 1451 (vw), 1383 (w), 1333 (m), 1268 (w), 1163 (w), 1076 (m), 1032(m, NO₂), 966 (vw), 711 (vw), 675 (vw), 587 (vw) cm⁻¹.

Synthesis of Product 71(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-({1-[3-(6-nitro-1H-indol-3-yl)propyl]-1H-1,2,3-triazol-4-yl}methyl)cyclohexanecarboxamide

Following general procedure III, alkyne 20 (14.4 mg, 18.5 μmol), azide54 (6.8 mg, 27.7 μmol), Na-L-ascorbate (186 μL, 18.6 μmol) andCuSO₄.5H₂O (92 μL, 9.20 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1(1 mL). After 1.5 h, triazole 71 (12.9 mg, 68%) was isolated bypreparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.47-0.71 (m, 4H, Cy), 0.82-0.94 (m, 1H, Cy),1.09 (d, J=6.4 Hz, 3H, Me), 1.12-1.44 (m, 11H, H-2_(a), H-6_(a), 5Cy,Lac H-3_(a)), 1.49 (ddd, J=3.6, 10.3, 13.6 Hz, 1H, Lac H-3_(b)),1.52-1.70 (m, 4H, (Cy, H-6_(b), H-5), 2.12-2.20 (m, 1H, H-2_(b)),2.26-2.36 (m, 3H, H-1, H-2″), 2.82 (t, J=7.5 Hz, 2H, H-3″), 3.13 (t,J=9.5 Hz, 1H, H-4), 3.53-3.58 (m, 2H, Gal H-5, Gal H-3), 3.65-3.81 (m,5H, H-3, Fuc H-2, Fuc H-4, Gal H-6), 3.84-3.90 (m, 2H, Fuc H-3, GalH-4), 3.94 (s, 1H, Lac H-2), 4.31-4.41 (m, 2H, H-1′), 4.46 (t, J=6.8 Hz,2H, H-1″), 4.65 (d, J=8.1 Hz, 1H, Gal H-1), 4.93-4.98 (m, 2H, Fuc H-1,Fuc H-5), 5.41 (dd, J=8.5, 9.3 Hz, 1H, Gal H-2), 7.30 (s, 1H, Ind H-2),7.38-7.44 (m, 2H, C₆H₅), 7.44-7.48 (m, 1H, Ind H-7), 7.55 (t, 1H, C₆H₅),7.82 (s, 1H, H-3′), 7.98-8.02 (m, 2H, C₆H₅), 8.02-8.07 (m, 1H, Ind H-6),8.45-8.48 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.72 (Fuc C-6),19.24 (Me), 22.56 (C3″), 26.53, 26.74, 27.33 (3C, Cy), 31.89 (C-2″),33.10, 34.29 (2C, Cy), 35.14, 35.20 (Cy, C-2), 35.63 (C-1′), 37.49(C-6), 39.33 (C-5), 43.04 (Lac C-3), 43.14 (C-1), 50.86 (C-1″), 62.92(Gal C-6), 67.71, 67.74 (2C, Gal C-4, Fuc C-5), 70.33 (Fuc C-2), 71.40(Fuc C-3), 72.99 (Gal C-2), 73.96 (Fuc C-4), 76.01 (Gal C-5), 79.76(C-3), 83.02 (C-4), 83.76 (Gal C-3), 100.41 (Fuc C-1), 100.55 (Gal C-1),112.53 (Ind C-7), 116.55 (Ind C-4), 117.68 (Ind C-3), 117.93 (Ind C-6),124.35 (C-3′), 127.11 (Ind C-2), 127.95 (Ind C-9), 129.66 (C₆H₅), 130.87(C₆H₅), 131.65 (C₆H₅), 134.24 (C₆H₅), 141.25 (Ind C-8), 142.29 (IndC-5), 166.84 (O(C═O)Ph), 176.98 (CONH); HR-MS: m/z calcd for C₅₀H₆₆N₆O₁₇[M+Na]⁺: 1045.4377. found: 1045.4381. [a]_(D) ²⁰=−54.1 (c=0.3, MeOH);HPLC (λ=350 nm): purity=95%, t_(R)=13.883 min; IR (KBr): ν=3433 (vs,OH), 2925 (m), 2852 (w), 1722 (m, C═O), 1631 (m, C═O), 1520 (w), 1468(w), 1451 (w), 1383 (w), 1333 (m, NO₂), 1270 (m), 1210 (w), 1163 (w),1108 (m), 1074 (m), 1029 (m), 966 (vw), 892 (vw), 807 (vw), 776 (vw),738 (vw), 712 (w), 672 (vw) cm⁻¹.

Synthesis of Product 72(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(2-{1-[2-(6-nitro-1H-indol-3-yl)ethyl]-1H-1,2,3-triazol-4-yl}ethyl)cyclohexanecarboxamide

Following general procedure III, alkyne 21 (15.2 mg, 19.2 μmol), azide53 (5.9 mg, 25.5 μmol), Na-L-ascorbate (96 μL, 9.60 μmol), andCuSO₄.5H₂O (48 μL, 4.80 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1(1 mL). After 3 h, triazole 72 (13.7 mg, 70%) was isolated bypreparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.50-0.73 (m, 4H, Cy), 0.84-0.96 (m, 1H, Cy),1.08 (d, J=6.5 Hz, 3H, Me), 1.10-1.26 (m, 3H, H-6_(a), H-2_(a, Cy),)1.26-1.37 (m, 8H, 5Cy, Fuc H-6), 1.37-1.44 (m, 1H, Lac H-3_(a)),1.46-1.58 (m, 3H, Lac H-3_(b), Cy, H-6_(b)), 1.59-1.68 (m, 1H, H-5),2.08-2.14 (m, 1H, H-2), 2.16-2.24 (m, 1H, H-1), 2.72 (t, J=7.2 Hz, 2H,H-2′), 3.12 (t, J=9.6 Hz, 1H, H-4), 3.24-3.29 (m, 2H, H-1′), 3.41 (t,J=6.7 Hz, 2H, H-2″), 3.54 (t, J=5.9 Hz, 1H, Gal H-5), 3.61 (dd, J=3.0,9.7 Hz, 1H, Gal H-3), 3.67 (ddd, J=4.7, 9.3, 11.6 Hz, 1H, H-3),3.70-3.81 (m, 4H, Fuc H-2, Fuc H-4, 2Gal H-6), 3.86 (dd, J=3.3, 10.3 Hz,1H, Fuc H-3), 3.94 (d, J=2.4 Hz, 1H, Gal H-4), 4.01 (dd, J=3.1, 9.7 Hz,1H, Lac H-2), 4.64-4.69 (m, 3H, Gal H-1, H-1″), 4.93-4.99 (m, 2H, FucH-1, Fuc H-5), 5.42 (dd, J=8.2, 9.6 Hz, 1H, Gal H-2), 7.22 (s, 1H, IndH-2), 7.41-7.48 (m, 3H, Ind H-7, C₆H₅), 7.51-7.57 (m, 1H, C₆H₅), 7.60(s, 1H, H-4′), 7.99-8.05 (m, 3H, Ind H-6, C₆H₅), 8.09 (s, 1H, NH),8.28-8.32 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.71 (Fuc C-6),19.24 (Me), 26.47, 26.53, 26.70 (3C, 2Cy, C-2′), 27.12, 27.26 (2C, Cy,C-2″), 33.14 (Cy), 34.18 (Cy), 35.04 (Cy), 35.29 (C-2), 37.43 (C-6),39.31 (C-5), 39.96 (C-1′, 42.77 (Lac C-3), 43.22 (C-1), 52.34 (C-1″),62.76 (Gal C-6), 67.71, 67.77 (2C, Gal C-4, Fuc C-5), 70.32 (Fuc C-2),71.42 (Fuc C-3), 73.02 (Gal C-2), 73.96 (Fuc C-4), 75.97 (Gal C-5),78.06 (Lac C-2), 79.83 (C-3), 83.01 (C-4), 83.67 (Gal C-3), 100.42 (FucC-1), 100.53 (Gal C-1), 112.58 (Ind C-7), 114.82 (Ind C-3), 116.36 (IndC-4), 117.99 (Ind C-6), 124.01 (C-4′), 127.93, 127.95 (2C, Ind C-2, IndC-9), 129.73 (C₆H₅), 130.88 (C₆H₅), 131.55 (C₆H₅), 134.37 (C₆H₅), 140.99(Ind C-8), 142.53 (Ind C-5), 146.19 (C-3′), 166.83 (O(C═O)Ph), 176.99(CONH), 178.96 (COOH); HR-MS: m/z calcd for C₅₀H₆₆N₆O₁₇ [M+Na]⁺:1045.4377. found: 1045.4375. [a]_(D) ²⁰=−53.9 (c=1.04, MeOH); HPLC(λ=350 nm): purity=100%, t_(R)=13.550 min; IR (KBr): ν=3431 (vs, OH),2927 (m), 2852 (w), 1721 (m, C═O), 1647 (m, C═O), 1545 (w), 1522 (w),1471 (w), 1450 (w), 1380 (w), 1334 (s, NO₂), 1272 (m), 1221 (w), 1163(w), 1097 (s), 1079 (s), 1029 (m), 999 (w), 963 (vw), 809 (vw), 785(vw), 741 (vw), 712 (w), 678 (w) cm⁻¹.

Synthesis of Product 73(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(2-{1-[3-(6-nitro-1H-indol-3-yl)propyl]-1H-1,2,3-triazol-4-yl}ethyl)cyclohexanecarboxamide

Following general procedure III, alkyne 21 (15.8 mg, 20.0 μmol), azide54 (6.9 mg, 28.1 μmol), Na-L-ascorbate (100 μL, 10.0 μmol), andCuSO₄.5H₂O (50 μL, 5.00 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1(1 mL). After 3 h, triazole 73 (14.1 mg, 68%) was isolated bypreparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.50-0.72 (m, 4H, Cy), 0.83-0.95 (m, 1H, Cy),1.00 (d, J=6.5 Hz, 3H, Me), 1.04-1.25 (m, 3H, H-2_(a), H-6_(a), Cy),1.26-1.36 (m, 7H, 4Cy, Fuc H-6), 1.36-1.44 (m, 1H, Lac H-3_(a)),1.45-1.62 (m, 4H, Lac H-3_(b), Cy, H-6_(b), H-5), 2.08-2.15 (m, 1H,H-2_(b)), 2.20 (tt, J=3.2, 12.7 Hz, 1H, H-1), 2.24-2.34 (m, 2H, H-2″),2.77-2.85 (m, 4H, H-2′, H-3″), 3.06 (t, J=9.6 Hz, 1H, H-4), 3.39 (t,J=7.0 Hz, 2H, H-1′), 3.52 (t, J=6.0 Hz, 1H, Gal H-5), 3.59 (dd, J=2.8,9.7 Hz, 1H, Gal H-3), 3.64 (ddd, J=4.6, 9.1, 11.6 Hz, 1H, H-3),3.69-3.81 (m, 4H, Fuc H-2, Fuc H-4, Gal H-6), 3.85 (dd, J=3.3, 10.3 Hz,1H, Fuc H-3), 3.93 (d, J=2.1 Hz, 1H, Gal H-4), 3.98 (dd, J=2.5, 9.6 Hz,1H, Lac H-2), 4.36-4.47 (m, 2H, H-1″), 4.64 (d, J=8.1 Hz, 1H, Gal H-1),4.90-4.93 (m, 1H, Fuc H-1), 4.96 (q, J=6.4 Hz, 1H, Fuc H-5), 5.42 (dd,J=8.2, 9.6 Hz, 1H, Gal H-2), 7.30 (s, 1H, Ind H-2), 7.43-7.50 (m, 3H,Ind H-7, C₆H₆), 7.55-7.61 (m, 1H, C₆H₆), 7.70 (s, 1H, H-4′), 8.01-8.09(m, 3H, C₆H₆, Ind H-6), 8.47-8.51 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz,MeOD): δ 16.73 (Fuc C-6), 19.17 (Me), 22.58 (C-3″), 26.53, 26.55, 26.71(3C, 2Cy, C-2′), 27.29 (Cy), 31.85 (C-2″), 33.12 (Cy), 34.24 (Cy), 35.12(Cy), 35.30 (C-2), 37.43 (C-6), 39.25 (C-5), 39.87 (C-1′), 42.95 (LacC-3), 43.17 (C-1), 50.75 (C-1″), 62.84 (Gal C-6), 67.72, 67.73 (2C, GalC-4, Fuc C-5), 70.31 (Fuc C-2), 71.41 (Fuc C-3), 73.03 (Gal C-2), 73.95(Fuc C-4), 75.97 (Gal C-5), 78.36 (Lac C-2)‡, 79.77 (C-3), 83.01 (C-4),83.72 (Gal C-3), 100.39 (Fuc C-1), 100.54 (Gal C-1), 112.53 (Ind C-7),116.59 (Ind C-4), 117.69 (Ind C-3), 117.93 (Ind C-6), 123.79 (C-4′),127.12 (Ind C-2), 127.94 (Ind C-9), 129.74 (C₆H₅), 130.91 (C₆H₅), 131.63(C₆H₅), 134.36 (C₆H₅), 141.23 (Ind C-8), 142.30 (Ind C-5), 146.29(C-3′), 166.84 (O(C═O)Ph), 177.00 (CONH); HR-MS: m/z calcd forC₅₁H₆₈N₆O₁₇ [M+H]⁺: 1037.4714. found: 1037.4714. [a]_(D) ²⁰=−59.7(c=1.22, MeOH); HPLC (λ=350 nm): purity=100%, t_(R)=13.717 min; IR(KBr): ν=3430 (vs, OH), 2927 (m), 2852 (w), 1720 (m, C═O), 1647 (m,C═O), 1547 (w), 1520 (w), 1471 (w), 1450 (w), 1333 (s, NO₂), 1272 (m),1221 (w), 1163 (w), 1097 (s), 1078 (s), 1029 (m), 999 (w), 966 (vw), 809(vw), 778 (vw), 738 (vw), 712 (w), 678 (vw) cm⁻¹.

Synthesis of Product 74 (1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(3-{1-[2-(6-nitro-1H-indol-3-yl)ethyl]-1H-1,2,3-triazol-4-yl}propyl)cyclohexanecarboxamide

Following general procedure III, alkyne 22 (14.5 mg, 18.0 μmol), azide53 (6.3 mg, 27.2 μmol), Na-L-ascorbate (90 μL, 9.00 μmol), andCuSO₄.5H₂O (45 μL, 4.50 μmol) were dissolved in ^(t)BuOH/H₂O/MeCN 1:1:1(1 mL). After 15 h, triazole 74 (10.3 mg, 55%) was isolated bypreparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.46-0.71 (m, 4H, Cy), 0.81-0.94 (m, 1H, Cy),1.11 (d, J=6.5 Hz, 3H, Me), 1.14-1.37 (m, 10H, H-2_(a), H-6_(a), 5Cy,Fuc H-6), 1.36-1.45 (m, 1H, Lac H-3_(a)), 1.45-1.56 (m, 2H, LacH-3_(b, Cy),) 1.56-1.63 (m, 1H, H-6_(b)), 1.62-1.74 (m, 3H, H-5, H-2′),2.11-2.19 (m, 1H, H-2_(b)), 2.20-2.30 (m, 1H, H-1), 2.53-2.64 (m, 2H,H-3′), 2.95-3.11 (m, 2H, H-1′), 3.15 (t, J=9.4 Hz, 1H, H-4), 3.41 (t,J=6.5 Hz, 2H, H-2″), 3.55-3.65 (m, 2H, Gal H-3, Gal H-5), 3.66-3.83 (m,5H, H-3, Fuc H-2, Fuc H-4, Gal H-6), 3.87 (dd, J=2.8, 10.3 Hz, 1H, FucH-3), 3.90-4.04 (m, 2H, Gal H-4 Lac H-2)), 4.61-4.74 (m, 3H, Gal H-1,H-1″), 4.95-5.01 (m, 2H, Fuc H-1, Fuc H-5), 5.38-5.48 (m, 1H, Gal H-2),7.23 (s, 1H Ind H-2,), 7.35-7.45 (m, 3H, Ind H-7, C₆H₅), 7.48-7.53 (m,1H, C₆H₅), 7.58 (s, 1H, H-5′), 7.99-8.05 (m, 3H, Ind H-6, C₆H₅),8.21-8.25 (m, 1H, Ind H-4); ¹³C-NMR (125 MHz, MeOD): δ 16.72 (Fuc C-6),19.28 (Me), 23.50 (C-3′), 26.50, 26.71 (2C, Cy), 27.09 (C-2″), 27.28(Cy), 30.12 (C-2′), 33.12, 34.38, 35.13 (3C, Cy), 35.41 (C-2), 37.45(C-6), 39.34, 39.40 (2C, C-1′, C-5), 43.33 (2C, Lac C-3, C-1), 52.54(C-1″), 62.88 (Gal C-5)), 67.73 (2C, Fuc C-5, Gal C-4), 70.33 (Fuc C-2),71.41 (Fuc C-3), 73.08 (Gal C-2), 73.97 (Fuc C-4), 76.26 (Lac C-2),79.84 (C-3), 83.03 (C-4), 83.70 (Gal C-3), 100.43 (Fuc C-1), 100.59 (GalC-1), 112.55 (Ind C-7), 114.91 (Ind C-3), 116.35 (Ind C-4), 117.93 (IndC-6), 123.98 (C-5′)‡, 127.99 (2C, Ind C-2, Ind C-9), 129.66 (C₆H₅),130.85 (C₆H₅), 131.59 (C₆H₅), 134.27 (C₆H₅), 140.96 (Ind C-8), 142.49(Ind C-5), 148.09 (C-4′)§, 166.82 (O(C═O)Ph), 177.03 (CONH); HR-MS: m/zcalcd for C₅₁H₆₈N₆O₁₇ [M+Na]⁺: 1059.4533. found: 1059.4528. [a]_(D)²⁰=−32.3 (c=0.31, MeOH); HPLC (λ=350 nm): purity=95%, t_(R)=13.617 min;IR (KBr): ν=3434 (vs, OH), 2926 (m), 282 (w), 1720 (w, C═O), 1631 (m,C═O), 1547 (w), 1520 (w), 1468 (w), 1449 (w), 1380 (w), 1333 (m, NO₂),1268 (m), 1169 (w), 1161 (w), 1097 (m), 1074 (m), 1032 (m), 999 (w), 712(w) cm⁻¹.

Synthesis of Product 75(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(3-{1-[3-(6-nitro-1H-indol-3-yl)propyl]-1H-1,2,3-triazol-4-yl}propyl)cyclohexanecarboxamide

Following general procedure III, alkyne 22 (11.6 mg, 14.4 μmol), azide54 (5.3 mg, 21.6), Na-L-ascorbate (72 μL, 7.20 μmol), and CuSO₄.5H₂O (36μL, 3.60 μmol) were dissolved in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL). After 2h, triazole 75 (9.5 mg, 63%) was isolated by preparative HPLC-MS.

¹H-NMR (500 MHz, MeOD): δ 0.46-0.67 (m, 4H, Cy), 0.85 (d, J=11.4 Hz, 1H,Cy), 1.10 (d, J=6.4 Hz, 3H, Me), 1.13-1.44 (m, 11H, H-2_(a), H-6_(a),5Cy, Fuc H-6, Lac H-3_(a)), 1.44-1.55 (m, 2H, Lac H-3_(b)), 1.53-1.62(m, 1H, H-6), 1.63 (d, J=16.3 Hz, 1H, H-5), 1.75-1.84 (m, 2H, H-2′),2.11-2.19 (m, 1H, H-2), 2.25 (tt, J=3.5, 13.0 Hz, 1H, H-1), 2.29-2.39(m, 2H, H-2″), 2.66-2.72 (m, 2H, H-3′), 2.82 (t, J=7.4 Hz, 2H, H-3″),3.06-3.23 (m, 3H, H-1′, H-4), 3.60 (dt, J=8.4, 11.3 Hz, 2H, Gal H-5, GalH-3), 3.65-3.84 (m, 5H, H-3, Fuc H-3, Fuc H-4, Gal H-6), 3.87 (dd,J=3.2, 10.3 Hz, 1H, Fuc H-3), 3.96 (s, 2H, Lac H-2, Gal H-4), 4.45 (t,J=6.8 Hz, 2H, H-1″), 4.69 (d, J=8.0 Hz, 1H, Gal H-1), 4.97 (ddd, J=4.9,14.3, 19.5 Hz, 2H, Fuc H-1, Fuc H-5), 5.38-5.45 (m, 1H, Gal H-2), 7.30(s, 1H, Ind H-2), 7.34-7.38 (m, 2H, C₆H₅), 7.44-7.46 (m, 1H, Ind H-7),7.48-7.52 (m, 1H, C₆H₅), 7.74 (s, 1H, H-5′), 7.98-8.02 (m, 2H, C₆H₅),8.02-8.05 (m, 1H, Ind H-6), 8.45-8.48 (m, 1H, Ind H-4); ¹³C-NMR (125MHz, MeOD): δ 16.72 (Fuc C-6), 19.25 (Me), 22.55 (C-3″), 23.63 (C-3′),26.46, 26.67, 27.25 (3C, Cy), 30.31 (C-2′), 31.86 (C-2″), 33.08, 34.19,35.07 (3C, Cy), 35.47 (C-2), 37.41 (C-6), 39.32 (C-5), 39.54 (C-1′),42.88 (Lac C-3), 43.36 (C-1), 50.80 (C-1″), 62.82 (Gal C-6), 67.71,67.73 (2C, Gal C-4, Fuc C-5), 70.34 (Fuc C-2), 71.42 (Fuc C-3), 72.99(Gal C-2), 73.97 (Fuc C-4), 75.99 (Gal C-5), 78, 69 (Lac C-2)‡, 79.89(C-3), 83.03 (C-4), 83.70 (Gal C-3), 100.44 (Fuc C-1), 100.58 (Gal C-1),112.51 (Ind C-7), 116.55 (Ind C-4), 117.69 (Ind C-3), 117.89 (Ind C-6),123.54 (C-5′), 127.11 (Ind C-2), 127.96 (Ind C-9), 129.64 (C₆H₅), 130.83(C₆H₅), 131.55 (C₆H₅), 134.27 (C₆H₅), 141.22 (Ind C-8), 142.25 (IndC-5), 148.53 (C-4′)§, 166.79 (O(C═O)Ph), 177.05 (CONH); HR-MS: m/z calcdfor C₅₂H₇₀N₆O₁₇ [M+H]⁺: 1051.4870. found: 1051.4870. [a]_(D) ²⁰=−47.6(c=0.87, MeOH); HPLC (λ=350 nm): purity=98%, t_(R)=13.767 min; IR (KBr):ν=3431 (vs, OH), 2927 (m), 2852 (w), 1720 (s, C═O), 1631 (m, C═O), 1547(w), 1517 (w), 1471 (w), 1450 (w), 1383 (w), 1332 (s, NO₂), 1270 (m),1213 (w), 1166 (w), 1106 (s), 1073 (s), 1032 (m), 996 (w), 963 (vw), 807(vw), 774 (vw), 741 (vw), 712 (w), 669 (vw) cm⁻¹.

Synthesis of Product 76(1R,3R,4R,5S)—N-(3-{1-[3-(5-Amino-1H-indol-1-yl)propyl]-1H-1,2,3-triazol-4-yl}propyl)-3-[2-O-benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methylcyclohexanecarboxamide

A mixture of nitroindole 66 (8.5 mg, 8.09 μmol), PtO₂ (2 mg, cat.),morpholine (5 μL) in MeOH was stirred at r.t. under an H₂ atmosphere(atm. pressure). Completion of the reaction was indicated bydiscoloration of the solution and confirmed by MS after 30 min. Thecatalyst was removed via filtration through a PTFE membrane filter, andthe solution was concentrated in vacuo. Purification by preparativeHPLC-MS (H₂O/MeCN+0.1% HCOOH) gave the amine 76 as a colorless solid(4.5 mg, 54%).

¹H-NMR (500 MHz, MeOD): δ 0.39-0.65 (m, 4H, Cy), 0.77-0.87 (m, 1H, Cy),1.04 (d, J=5.8 Hz, 3H, Me), 1.09-1.36 (m, 11H, H-2_(a), H-6_(a), 5Cy,Fuc H-6, Lac H-3), 1.43 (t, J=11.9 Hz, 1H, Lac H-3), 1.47-1.55 (m, 2H,H-6, Cy), 1.55-1.64 (m, 1H, H-5), 1.70-1.78 (m, 2H, H-2′)), 2.05-2.12(m, 1H, H-2), 2.19 (t, J=11.8 Hz, 1H, H-1), 2.39 (p, J=6.6 Hz, 2H,H-2″), 2.62 (t, J=7.0 Hz, 2H, H-3′), 3.02-3.16 (m, 3H, H-1′, H-4),3.47-3.54 (m, 2H, Gal H-3, Gal H-3, Gal H-5), 3.60-3.80 (m, 6H, H-3, FucH-2, Fuc H-4, Gal H-6, Lac H-2), 3.80-3.86 (m, 1H, Fuc H-3), 3.89 (s,1H, Gal H-4), 4.20 (t, J=6.5 Hz, 2H, H-3″), 4.30 (t, J=6.3 Hz, 2H,H-1″), 4.62 (d, J=8.1 Hz, 1H, Gal H-1), 4.89-4.97 (m, 2H, Fuc H-1, FucH-5), 5.37 (t, J=8.7 Hz, 1H, Gal H-2), 6.41 (s, 1H, Ind H-3), 6.92-6.98(m, 1H, Ind H-6), 7.27 (s, 1H, Ind H-2), 7.29-7.37 (m, 4H, C₆H₅, IndH-4, Ind H-7), 7.42-7.48 (m, 1H, C₆H₅), 7.60 (s, 1H, H-5′), 7.94-8.00(m, 2H, C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 16.75 (Fuc C-6), 19.26 (Me),23.54 (C-3′), 26.53, 26.77, 27.34 (3C, Cy), 30.15 (C-2′), 31.60 (C-2″),33.05, 34.33, 35.26 (3C, Cy), 35.50 (C-2), 37.39 (C-6), 39.26 (C-5),39.41 (C-1′), 43.27 (2C, Lac C-3, C-1), 44.28 (C-3″), [48.66 (C-1″)],63.02 (Gal C-6), 67.70, 67.78 (2C, Gal C-4, Fuc C-5), 70.28 (Fuc C-2),71.36 (Fuc C-3), 73.07 (Gal C-2), 73.94 (Fuc C-4), 76.10 (Gal C-5),79.74 (C-3), 79.89 (Lac C-2), 83.02 (C-4), 83.66 (Gal C-3), 100.46 (FucC-1), 100.62 (Gal C-1), 102.22 (Ind C-3), 111.54 (Ind C-4), 112.92 (IndC-6), 115.87 (Ind C-7), 123.58 (C-5′), 129.63 (C₆H₅), 130.84, 130.87(2C, C₆H₅, Ind H-2), 131.68 (Ind C-8), 134.19 (C₆H₅), 135.30 (Ind C-5),148.45 (C-4′), 166.78 (O(C═O)Ph), 177.05 (CONH), 181.91 (COOH); [a]_(D)²⁰=−59.1 (c=0.45, MeOH); HPLC (λ=350 nm): purity=97%, t_(R)=11.167 min;IR (KBr): ν=3431 (vs, OH), 2926 (m), 2853 (w), 1722 (w), 1643 (m), 1603(m), 1584 (m), 1555 (w), 1492 (w), 1451 (w), 1406 (w), 1384 (w), 1365(w), 1348 (w), 1316 (w), 1273 (m), 1222 (vw), 1167 (w), 1118 (m), 1096(m), 1079 (s), 1031 (m), 1000 (w), 967 (vw), 804 (vw), 768 (vw), 713 (m)cm⁻¹.

Synthesis of Product 77(1R,3R,4R,5S)—N-(3-{1-[3-(5-Acetamido-1H-indol-1-yl)propyl]-1H-1,2,3-triazol-4-yl}propyl)-3-[2-O-benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methylcyclohexanecarboxamide

Following general procedure III, alkyne 22 (11.6 mg, 14.4 μmol), azide38 (13 mg, 22.3 μmol), Na-L-ascorbate (72 μL, 7.20 μmol) and CuSO₄.5H₂O(36 μL, 3.60 μmol) were stirred in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL; noprior degassing). After removal of the solvent under reduced pressure,the crude product was redissolved in MeOH, and centrifuged. Thesupernatant was subjected to preparative HPLC-MS to afford 77 as acolorless solid (11.0 mg, 73%).

¹H-NMR (500 MHz, MeOD): δ 0.47-0.57 (m, 1H, Cy), 0.57-0.70 (m, 3H, Cy),0.82-0.94 (m, 1H, Cy), 1.10 (d, J=6.4 Hz, 3H, Me), 1.13-1.36 (m, 8H,H-2_(a), H-6_(a), 5Cy, Fuc H-6), 1.35-1.44 (m, 1H, Lac H-3_(a)),1.44-1.51 (m, 1H, Lac H-3_(b)), 1.51-1.61 (m, 2H, H-6_(b), Cy),1.61-1.70 (m, 1H, H-5), 1.73-1.80 (m, 2H, H-2′), 2.10-2.15 (m, 4H,H-2_(b), NHAc-Me), 2.21-2.29 (m, 1H, H-1), 2.42 (p, J=6.7 Hz, 2H, H-2″),2.65 (t, J=7.5 Hz, 2H, H-3′), 3.06-3.18 (m, 3H, H-1′, H-4), 3.53-3.57(m, 1H, Gal H-5), 3.60 (dd, J=2.8, 9.7 Hz, 1H, Gal H-3), 3.65-3.81 (m,5H, H-3, Fuc H-2, Fuc H-4, Gal H-6), 3.86 (dd, J=3.2, 10.3 Hz, 1H, FucH-3), 3.94 (d, J=2.1 Hz, 1H, Gal H-4), 4.00 (dd, J=2.3, 9.6 Hz, 1H, LacH-2), 4.20 (t, J=6.7 Hz, 2H, H-3″), 4.32 (t, J=6.8 Hz, 2H, H-1″), 4.67(d, J=8.0 Hz, 1H, Gal H-1), 4.94-5.01 (m, 2H, Fuc H-1, Fuc H-5),5.40-5.44 (m, 1H, Gal H-2), 6.40-6.43 (m, 1H, Ind H-3), 7.18-7.28 (m,3H, Ind H-2, Ind H-6, Ind H-7), 7.33-7.41 (m, 2H, C₆H₅), 7.47-7.52 (m,1H, C₆H₅), 7.60 (s, 1H, H-5′), 7.75-7.79 (m, 1H, Ind H-4), 7.99-8.03 (m,2H, C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 16.74 (Fuc C-6), 19.25 (Me), 23.54(Ac-Me), 23.67 (C-3′), 26.50, 26.68, 27.25 (3C, Cy), 30.23 (C-2′), 31.61(C-2″), 33.07, 34.14, 35.05 (3C, Cy), 35.43 (C-2), 37.43 (C-6), 39.33,39.47 (2C, C-5, C-1′), 42.81 (Lac C-3), 43.30 (C-1), 44.14 (C-3″), 48.66(C-1″)‡, 62.76 (Gal C-6), 67.70 (2C, Gal C-4, Fuc C-5), 70.28 (Fuc C-2),71.39 (Fuc C-3), 72.95 (Gal C-2), 73.96 (Fuc C-4), 75.95 (Gal C-5),78.15 (Lac C-2), 79.80 (C-3), 83.03 (C-4), 83.65 (Gal C-3), 100.47 (FucC-1), 100.50 (Gal C-1), 102.48 (Ind C-3), 110.34 (Ind C-7), 114.12 (IndC-4), 117.20 (Ind C-6), 123.59 (C-5′), 129.68 (C₆H₅), 129.97 (Ind C-9),130.11 (Ind C-2), 130.83 (C₆H₅), 131.52, 131.94 (2C, Ind C-5, C₆H₅),134.32 (C₆H₅), 134.78 (Ind C-8), 147.95 (C-4′), 166.76 (O(C═O)Ph),171.50 (MeCONH), 177.04 (CONH), 178.70 (COOH). [a]_(D) ²⁰=−49.4 (c=0.83,MeOH); HPLC (λ=350 nm): purity=95%, t_(R)=12.483 min; IR (KBr): ν=3422(vs, OH), 2927 (s), 2853 (m), 1725 (s, C═O), 1651 (s, C═O), 1603 (m),1587 (m), 1548 (m), 1488 (m), 1450 (m), 1401 (w), 1373 (m), 1337 (m),1316 (m), 1271 (vs), 1113 (vs), 1076 (vs), 1033 (s), 1000 (w), 874 (vw),804 (w), 762 (w), 762 (w), 713 (m), 675 (w) cm⁻¹.

Synthesis of Product 78(1R,3R,4R,5S)—N-(3-{1-[3-(5-{2-[2-(2-Aminoethoxy)ethoxy]acetamido}-1H-indol-1-yl)propyl]-1H-1,2,3-triazol-4-yl}propyl)-3-[2-O-benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methylcyclohexanecarboxamide

Following general procedure III, alkyne 22 (12.0 mg, 14.9 μmol), azide40 (13 mg, 22.3 μmol), Na-L-ascorbate (75 μL, 7.50 μmol) and CuSO₄.5H₂O(38 μL, 3.80 μmol) were stirred in ^(t)BuOH/H₂O/THF 1:1:1 (1 mL; noprior degassing) for 2 h, followed by the addition of piperidine (4.4μL). After another hour, further 10 μL of piperidine was added, andstirring was continued for 2 h. Following removal of the solvent invacuo, the residue was dissolved in MeOH, filtrated through a PTFEmembrane filter, and the solution was centrifuged. The supernatant wassubjected to preparative HPLC-MS to afford 78 as a colorless solid (11.4mg, 66%).

¹H-NMR (500 MHz, MeOD): δ 0.39-0.69 (m, 4H, Cy), 0.79-0.93 (m, 1H, Cy),1.09 (d, J=6.4 Hz, 3H, Me), 1.12-1.41 (m, 11H, H-2_(a), H-6_(a), 4Cy,Fuc H-6, Lac H-3_(a)), 1.41-1.51 (m, 1H, Lac H-3), 1.51-1.61 (m, 2H,H-6_(b, Cy),) 1.61-1.73 (m, 1H, H-5), 1.73-1.84 (m, 2H, H-2′), 2.10-2.15(m, 1H, H-2_(b)), 2.21-2.29 (m, 1H, H-1), 2.44 (p, J=6.6 Hz, 2H, H-2″),2.62-2.69 (m, 2H, H-3′), 3.05-3.20 (m, 5H, H-1′, H-4, H-6), 3.49 (d,J=9.2 Hz, 1H, Gal H-3), 3.52-3.57 (m, 1H, Gal H-5), 3.65-3.83 (m, 12H,H-3, Fuc H-2, Fuc H-4, Gal H-6, H-3*, H-4*, H-5*, Lac H-2), 3.86 (dd,J=3.2, 10.3 Hz, 1H, Fuc H-3), 3.89-3.94 (m, 1H, Gal H-4, 4.18-4.25 (m,4H, H-2*, H-3″), 4.34 (t, J=6.7 Hz, 2H, H-1″), 4.64 (d, J=8.1 Hz, 1H,Gal H-1), 4.94-5.00 (m, 3H, Fuc H-1, Fuc H-5), 5.39 (t, J=8.8 Hz, 1H,Gal H-2), 6.43-6.46 (m, 1H, Ind H-3), 7.24-7.27 (m, 1H, Ind H-2), 7.28(s, 2H, Ind H-6, Ind H-7), 7.31-7.36 (m, 2H, C₆H₅), 7.43-7.49 (m, 1H,C₆H₅), 7.61 (s, 1H, H-5′), 7.83 (s, 1H, Ind H-4), 7.95-8.01 (m, 2H,C₆H₅); ¹³C-NMR (125 MHz, MeOD): δ 16.79 (Fuc C-6), 19.29 (Me), 23.54(C-3′), 26.52, 26.77, 27.36 (3C, Cy), 30.19 (C-2′), 31.60 (C-2″), 33.03,34.37, 35.33 (3C, Cy), 35.53 (C-2), 37.40 (C-6), 39.25, 39.42 (2C, C-5,C-1′), 40.57 (C-6*), 43.23 (C-1), 43.43 (Lac C-3), 44.20 (C-3″), 48.69(C-1″)‡, 63.06 (Gal C-6), 67.59, 67.74 (2C, Gal C-4, Fuc C-5), 68.00(C-5*), 70.27 (Fuc C-2) 70.33, 71.36 (2C, Fuc C-3, C-2), 71.48 (2C,C-3*, C-4), 73.00 (Gal C-2), 73.94 (Fuc C-4), 76.06 (Gal C-5), 79.75(C-3), 83.02 (C-4), 83.69 (Gal C-3), 100.39 (Fuc C-1), 100.62 (Gal C-1),102.57 (Ind C-3), 110.52 (Ind C-7), 114.43 (Ind C-), 117.31 (Ind C-6),123.66 (C-5′), 129.62 (C₆H₅), 130.13, 130.18 (2C, Ind C-2, Ind C-9),130.82 (C₆H₅), 130.96, 131.66 (2C, Ind C-5, C₆H₅), 134.16 (C₆H₅), 135.01(Ind C-8), 148.82 (C-4′), 166.78 (O(C═O)Ph), 170.59 (C-1*), 177.03(CONH); HPLC (λ=350 nm): purity=95%, t_(R)=12.483 min; [a]_(D) ²⁰=−44.4(c=0.67, MeOH); IR (KBr): ν=3430 (vs, OH), 3137 (m, NH₂), 2925 (vs),2855 (m), 1724 (m), 1720 (m), 1654 (s), 1649 (s), 1586 (s), 1542 (m),1489 (m), 1450 (m), 1401 (w), 1385 (w), 1340 (w), 1293 (w), 1272 (s),1219 (vw), 1163 (w), 1117 (vs), 1098 (vs), 1079 (vs), 1032 (s), 997 (w),967 (vw), 802 (vw), 763 (vw), 714 (w) cm⁻¹.

Example 4 Synthesis of the Selectin Antagonist with Piperazine Linker(FIG. 7) Synthesis of Intermediate 793-{4-[3-(5-Nitro-1H-indol-1-yl)propyl]piperazin-1-yl}propan-1-ol

Bromide 25 (176 mg, 0.622 mmol), 3-(piperazin-1-yl)propan-1-ol (278 mg,1.93 mmol), and Et₃N (259 μL, 1.87 mmol) were dissolved in anhydrous DMFand stirred at r.t. for 2 h. After removal of the solvent in vacuo, thecrude product was subjected to silica gel chromatography (MeOH inCH₂Cl₂, gradient 0 to 20 to 80%) to afford alcohol 79 (215 mg, quant.)as yellow crystals.

¹H-NMR (500 MHz, CDCl₃): δ 1.70-1.76 (m, 2H, H-2″), 1.99 (p, J=6.7 Hz,2H, H-2′), 2.04-3.07 (bs, 8H, Pip-H), 2.23 (t, J=6.7 Hz, 2H, H-3′),2.61-2.66 (m, 2H, H-1″), 3.77-3.81 (m, 2H, H-3″), 4.25 (t, J=6.6 Hz, 2H,H-1′), 6.64-6.68 (m, 1H, H-3), 7.26-7.28 (m, 1H, H-2), 7.39-7.44 (m, 1H,H-7), 8.06-8.10 (m, 1H, H-6), 8.54-8.57 (m, 1H, H-4); ¹³C-NMR (125 MHz,CDCl₃): δ 27.06 (C-2″), 27.12 (C-2′), 44.10 (C-1′), 52.92 (2C, C-a,C-a′), 53.28 (2C, C-b, C-b′), 54.24 (C-3′), 58.61 (C-1″), 64.38 (C-3″),103.99 (C-3), 109.44 (C-7), 117.01 (C-6), 118.18 (C-4), 127.58 (C-9),131.23 (C-2), 138.98 (C-8), 141.39 (C-5); IR (KBr): ν=3435 (s, OH), 2947(m), 2922 (m), 2826 (m), 1634 (vw), 1610 (w), 1578 (vw), 1513 (s), 1479(m), 1464 (m), 1404 (w), 1369 (w), 1328 (vs, NO₂), 1306 (s), 1274 (m),1186 (w), 1157 (m), 1130 (m), 1067 (s), 1006 (m) 899 (vw), 779 (w), 751(m), 718 (w) cm⁻¹.

Synthesis of Intermediate 801-{3-[4-(3-Azidopropyl)piperazin-1-yl]propyl}-5-nitro-1H-indole

Alcohol 79 (181 mg, 0.522 mmol) and diphenyl phosphoryl azide (DPPA; 134μL, 0.626 mmol) were dissolved in anhydrous THF (2 mL). The solution wascooled to −15° C. in a salt-ice bath, and diazobicyclo undecene (DBU;93.6 μL, 0.626 mmol) was added dropwise, and the solution was allowed towarm to r.t. After 2 h, NaN₃ (71 mg, 1.09 mmol) was added to thereaction mixture as there was no reaction progress according toTLC(CH₂Cl₂, MeOH 10:1). After 12 h of stirring, additional 0.5 eq. ofDPPA (56 μL, 0.261 mmol) and DBU (39 μL, 0.261 mmol) were added, andstirring was continued for 2 h. The solvent was removed under reducedpressure, the crude mixture was dissolved in EtOAc (30 mL), washed withsatd. aq. NaHCO₃ (20 mL) and brine (20 mL). The aqueous layers wereextracted with EtOAc (2×30 mL), and the organic phase was dried overNa₂SO₄. After removal of the solvent in vacuo, the mixture was purifiedusing silica gel chromatography (MeOH in CH₂Cl₂, gradient 0 to 15 to70%) to afford a mixture of azide 80 and its diphenyl phosphoryl ester(note: in TLC, azide and ester were not separable using CH₂Cl₂, MeOH10:1). The mixture of azide and ester was dissolved in anhydrous DMF,and NaN₃ (171 mg, 2.63 mmol) was added. The mixture was stirred at 60°C. for 25 h until TLC (on deactivated TLC plates; petroleum ether, EtOAc1:1+Et₃N) and MS indicated full conversion of the phosphoryl ester. H₂O(20 mL) was added, and the aqueous phase was extracted with EtOAc (3×50mL). The organic layers were washed with NaHCO₃ (20 mL) and brine (20mL) and subsequently dried over Na₂SO₄. After removing the solvent invacuo, the crude product was purified using silica gel chromatography(MeOH in CH₂Cl₂, gradient 0 to 10 to 70%) to give azide 80 (154 mg, 79%)as a viscous yellow oil.

¹H-NMR (500 MHz, CDCl₃): δ 1.74-1.81 (p, J=6.7 Hz, 2H, H-2″), 2.00 (p,J=6.7 Hz, 2H, H-2′), 2.23 (t, J=6.7 Hz, 2H, H-1′), 2.28-2.70 (m, 11H,Pip-H, H-1″), 3.34 (t, J=6.7 Hz, 2H, H-3″), 4.26 (t, J=6.6 Hz, 2H,H-1′), 6.65-6.68 (m, 1H, H-3), 7.25-7.29 (m, 1H, H-2), 7.41-7.45 (m, 1H,H-7), 8.07-8.11 (m, 1H, H-6), 8.56-8.59 (m, 1H, H-4); ¹³C-NMR (125 MHz,CDCl₃): δ 26.35 (C-2″), 27.27 (C-2′), 44.21 (C-1′), 49.65 (C-3″), 53.07,53.28 (4C, a/a′, b/b′), 54.40 (C-1′), 55.34 (C-1″), 104.05 (C-3), 109.53(C-7), 117.13 (C-6), 118.28 (C-4), 127.66 (C-9), 131.30 (C-2), 139.08(C-8), 141.52 (C5).

Synthesis of Intermediate 813-{4-[3-(5-Nitro-1H-indol-1-yl)propyl]piperazin-1-yl}propan-1-amine

Azide 80 (149 mg, 0.401 mmol) and triphenyl phosphine (129 mg, 0.492mmol) were dissolved in THF (4 mL) and H₂O (1 mL). The solution wasstirred at 60° C. for 3 h, and the solvent was removed under reducedpressure. The mixture was redissolved in MeOH and subjected topreparative HPLC-MS (H₂O/MeCN+0.2% HCOOH) to afford amine 81 (108 mg,78%) as a viscous yellow oil.

¹H-NMR (500 MHz, MeOD): δ 1.89 (p, J=7.0 Hz, 2H, H-2″), 2.14-2.21 (m,2H, H-2′), 2.65-2.72 (m, 4H, H-1″, H-3′), 2.83 (d, J=50.9 Hz, 8H, PipH-b/b′, Pip H-a-/a′), 3.01 (t, J=7.2 Hz, 2H, H-3″), 4.36 (t, J=6.8 Hz,2H, H-1′), 6.72-6.74 (m, 1H, H-3), 7.48-7.50 (m, 1H, H-2), 7.58-7.62 (m,1H, H-7), 8.06-8.09 (m, 1H, H-6), 8.53-8.55 (m, 1H, H-4); ¹³C-NMR (125MHz, MeOD): δ 24.46 (C-2″), 27.05 (C-2′), 39.21 (C-3′), 44.84 (C-1′,52.20 (C-b/b′), 52.70 (C-a/a′), 55.28 (C-3′), 55.67 (C-1″), 105.10(C-3), 110.80 (C-7), 117.81 (C-6), 118.79 (C-4), 129.29 (C-9), 132.83(C-2), 140.36 (C-8), 142.76 (C-5).

Synthesis of Product 82(1R,3R,4R,5S)-3-[2-O-Benzoyl-3-O-((1S)-1-carboxy-2-cyclohexyl-ethyl)-(β-D-galactopyranosyl)oxy]-4-[(α-L-fucopyranosyl)oxy]-5-methyl-N-(3-{1-[3-(5-nitro-1H-indol-1-yl)propyl]piperazin-1-yl}propyl)cyclohexanecarboxamide

Diacid 15 (30 mg, 40.5 μmol) and HOBt (17.0 mg, 126μmol) were dissolvedin anhydrous DMF under argon. At r.t., HBTU (18.4 mg, 48.5 μmol) wasadded. After 5 min of stirring, the solution was transferred via asyringe to a flask containing amine 81 (89 mg, 258 μmol) and DIPEA (0.3mL). The solvent was removed in vacuo after another 2.5 h of stirring,and the crude product was purified using preparative HPLC-MS(H₂O/MeCN+0.2% HCOOH). Product 82 (14.6 mg, 34%) was obtained as ayellow solid.

¹H-NMR (500 MHz, MeOD): δ 0.41-0.72 (m, 4H, Cy), 0.80-0.94 (m, 1H, Cy),1.10 (d, J=6.3 Hz, 3H, Me), 1.14-1.42 (m, 11H, H-6_(a), H-2_(a), 5Cy,Fuc H-6, Lac H-3_(a)), 1.45-1.53 (m, 1H, Lac H-3_(b)), 1.55-1.72 (m, 5H,Cy, H-6, H-2′, H-5), 2.00-2.09 (m, 2H, H-2″), 2.11-2.20 (m, 1H, H-2),2.20-2.70 (m, 12H, H-1, H-1″, H-3′, Pip-H), 3.13 (t, J=6.7 Hz, 2H,H-1′), 3.17 (t, J=9.6 Hz, 1H, H-4), 3.51-3.61 (m, 2H, Gal H-3, Gal H-5),3.67-3.83 (m, 6H, H-3, Lac H-2, Gal H-6, Fuc H-4, Fuc H-2), 3.89 (dd,J=3.1, 10.2 Hz, 1H, Fuc H-3), 3.91-3.96 (m, 1H, Gal H-4), 4.33 (t, J=6.6Hz, 2H, H-3″), 4.69 (d, J=8.1 Hz, 1H, Gal H-1), 4.94-5.04 (m, 2H, FucH-1, Fuc H-5), 5.42 (t, J=8.9 Hz, 1H, Gal H-2), 6.66-6.77 (m, 1H, IndH-3), 7.43-7.54 (m, 3H, C₆H₅, Ind H-2), 7.55-7.66 (m, 2H, C₆H₅, IndH-7), 8.00-8.13 (m, 3H, C₆H₅, Ind H-6), 8.52-8.59 (m, 1H, Ind H-4);¹³C-NMR (125 MHz, MeOD): δ 16.77 (Fuc C-6), 19.31 (Me), 26.55, 26.80,26.92 (4C, Cy, C-2′), 27.39 (C-1″), 28.17 (C-2″), 33.03, 34.42, 35.38(3C, Cy), 35.48 (C-2), 37.67 (C-6), 39.06, 39.31 (2C, C-5, C-1′), 43.45(C-1), 43.54 (Lac C-3), 45.30 (C-3″), 53.83, 53.92 (4C, Pip-C), 56.06(C-1″), 57.37 (C-3′), 63.15 (Gal C-6), 67.63, 67.75 (2C, Gal C-4, FucC-5), 70.28 (Fuc C-2), 71.34 (Fuc C-3), 73.03 (Gal C-2), 73.93 (FucC-4), 76.04 (Gal C-5), 79.69 (C-3), 80.65 (Lac C-2), 82.94 C-4), 83.79(Gal C-3), 100.36 (Fuc C-1), 100.64 (Gal C-1), 104.83 (Ind C-3), 110.94(Ind C-7), 117.64 (Ind C-6), 118.74 (Ind C-4), 129.20 (Ind C-9), 129.68(C₆H₅), 130.89 (C₆H₅), 131.81 (C₆H₅), 132.97 (Ind C-2), 134.21 (C₆H₅),140.47 (Ind C-8), 142.63 (Ind C-5), 166.81 (O(C═O)Ph), 176.97 (CONH),183.25 (COOH).

Example 5 Biological Evaluation BIAcore Assay

The biological evaluation of the E-selectin antagonists was performed bysurface plasmon resonance (SPR). For this purpose, an E-selectin/IgGconstruct was captured onto a BIAcore chip surface bearing covalentlylinked goat anti-human Fc specific antibody (Biacore Life Sciences,Uppsala, Sweden).

Ranking Procedure

The ranking of the antagonists was performed in order to obtainqualitative information on the binding strength of twenty indoleantagonists. For the ranking, SPR signals were recorded at a singleconcentration (0.05 μM) and divided by the molecular weight of thecompounds. The result obtained was normalized to the response obtainedwith an internal standard (63) in order to avoid problems related to themeasurement on different chips with different surfaces. DMSO calibrationand double referencing were applied to correct bulk effects and othersystematic artifacts (subtraction of reference surface and blankinjection).

Evaluation of E-Selectin Antagonists with Surface Plasmon Resonance(SPR)-Based Assay

Ten twofold serial dilutions of the ligand were non-randomly injected.The E-selectin antagonists were diluted in running buffer (HBS-Psupplemented with 20 mm calcium and DMSO 5% v/v). Antagonist dilutionswere injected with a 600 s association time and 600 s dissociation timeat a flow rate of 20 μL/min over the reference and the active flow cell.Saturation was reached at a concentration of 1 μm and a total of eleventwofold dilutions were injected. In addition, to prevent the presence ofresidual traces of compound, a blank injection was performed betweeneach injection of ligand dilution. DMSO calibration and doublereferencing were applied to correct bulk effects and other systematicartifacts (subtraction of reference surface and blank injection).

The results are summarized in Table 1.

TABLE 1 Absolute and relative* binding affinities K_(D) val- No.Structure ues 56

>89 nM* 57

>89 nM* 58

 57 nM 59

>89 nM* 60

>89 nM* 61

>89 nM* 62

>89 nM* 63

 49 nM 64

 89 nM 65

>89 nM* 66

 30 nM 67

>89 nM* 68

>89 nM* 69

>89 nM* 70

>89 nM* 71

>89 nM* 72

>89 nM* 73

 50 nM 74

>89 nM* 75

>89 nM* 76

>89 nM* 77

>89 nM* 78

>89 nM* 82

110 nM *Affinity determined in the ranking procedure described above,i.e., the K_(D) values are larger than for reference compound 64,however below 135 nM.

All of the above U.S. patents, U.S. patent application publications,U.S. patent applications, foreign patents, foreign patent applicationsand non-patent publications referred to in this specification and/orlisted in the Application Data Sheet, are incorporated herein byreference, in their entirety.

From the foregoing it will be appreciated that, although specificembodiments of the invention have been described herein for purposes ofillustration, various modifications may be made without deviating fromthe spirit and scope of the invention.

1. A compound or physiologically acceptable salt thereof, having one ofthe formulae:

2.-14. (canceled)
 15. A compound or physiologically acceptable saltthereof, having one of the formulae:

16.-22. (canceled)
 23. A compound or physiologically acceptable saltthereof, having one of the formulae:


24. (canceled)
 25. A composition comprising the compound or salt thereofaccording to claim 1 in combination with a pharmaceutically acceptablecarrier or diluent.
 26. The compound or salt thereof according to claim1 for inhibiting an E-selectin.
 27. A method for treating a disease orcomplication associated therewith in an individual wherein anE-selectin-mediated function is involved, said method comprisingadministering to the individual the compound of claim
 1. 28. The methodaccording to claim 27 wherein the E-selectin-mediated function is anE-selectin-mediated intercellular adhesion. 29.-31. (canceled)
 32. Acomposition comprising the compound or salt thereof according to claim15 in combination with a pharmaceutically acceptable carrier or diluent.33. The compound or salt thereof according to claim 15 for inhibiting anE-selectin.
 34. A method for treating a disease or complicationassociated therewith in an individual wherein an E-selectin-mediatedfunction is involved, said method comprising administering to theindividual the compound of claim
 15. 35. The method according to claim34 wherein the E-selectin-mediated function is an E-selectin-mediatedintercellular adhesion.
 36. A composition comprising the compound orsalt thereof according to claim 23 in combination with apharmaceutically acceptable carrier or diluent.
 37. The compound or saltthereof according to claim 23 for inhibiting an E-selectin.
 38. A methodfor treating a disease or complication associated therewith in anindividual wherein an E-selectin-mediated function is involved, saidmethod comprising administering to the individual the compound of claim23.
 39. The method according to claim 38 wherein the E-selectin-mediatedfunction is an E-selectin-mediated intercellular adhesion.